| Title of Invention | MONOCLONAL ANTIBODY REACTIVE TO HUMAN CHOLESTROL ESTER TRANSFER PROTEIN. |
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| Abstract | MONOCLONAL ANTIBODIES WHICH HAS BINDING SPECIFICITY TO THE HUMAN CETP (CETP INHIBITION ACTIVITY) AVAILABLE AS REAGENTS FOR THE HUMAN CETP PURIFICATION OR DETERMINATION, AND PHARMACEUTICALS OF PREVENTION AND/OR TREATMENT FOR HYPERLIPIDEMIA OR ARTERIOSCLEROSIS ARE PROVIDED. FURTHERMORE, THE PURIFICATION OF THE DETERMINATION METHODS OF THE HUMAN CETP BY USING THE MONOCLONAL ANTIBODY IS ALSO PROVIDED. |
| Full Text | FIELD OF THE INVENTION The present invention relates to hybridoma producing monoclonal antibody reactive to human cholesterol ester transfer protein(CETP), monoclonal antibody and its fragment reactive to human CETP, immobilized monoclonal antibody and immobilized antibody fragment, labeled monoclonal antibody and labeled antibody fragment, a kit for detection, assay, separation or purification of human CETP, a method for detection, assay, separation and purification of human CETP and a pharmaceutical composition containing said monoclonal antibody or said antibody fragment. This application is divided out of Indian application no.782/Cal/96. BACKGROUND OF THE INVENTION There are three types of cholesterol, free type, long chain fatty acid type and ester type, in all the tissues and blood plasma in organisms. The former two play important role in consisting of cell membrane. The latter is physiologically inactive and is mainly existing as a storage form. The cholesterols are derived from ingestion at small intestine or from biosynthesis at various tissues, especially at liver. Most of the cholesterols are derived from biosynthesis at liver. Free cholesterol biosynthesized and secreted from liver is incorporated in very low density lipoprotein(VLDL). Then, by the actions of lipoprotein lipase(LPL) and hepatic triglyceride lipase(HTGL), it is metabolized to low density 1A lipoprotein (LDL) through intermediate density lipoprotein (IDL). By incorporation of LDL into peripheral cells through LDL receptor, free cholesterol is supplied to cells. There is a pathway from peripheral cells to liver called cholesterol reverse transfer system which is reverse direction of the way from liver to peripheral cells as mentioned above. Surplus free cholesterol supplied in peripheral cells from liver is drawn by high density lipoprotein (HDL) in blood. Then, by the action of lecithin cholesterol acyl transferase (LCAT), it is converted to cholesterol ester and is stored in high density lipoprotein (HDL) in blood. By the action of cholesterol ester transfer protein(CETP), the cholesterol ester stored in HDL is transferred to VLDL, IDL or LDL in blood. By incorporation of VLDL, IDL or LDL having cholesterol ester into liver through LDL receptor, cholesterol is indirectly transferred to liver. Recently, the reverse cholesterol transfer system draws much attention as a mechanism to prevent atherosclerosis by preventing peripheral cells from accumulation of cholesterol. In fact, as for HDL which plays important role in the reverse cholesterol transfer system, many epidemiological surveys show that decrease of cholesterol ester in blood HDL is one of the risk factors of coronary artery disorders. It is now well recognized that HDL is a lipoprotein having anti atherosclerosis action. In addition to the importance of blood HDL, it becomes recognized that CETP is also important because it mediates transfer of cholesterol ester in HDL into blood LDL. Therefore, it becomes an urgent matter to elucidate relationship between CETP and various diseases such as CETP deficiency, hyperlipemia, hyperalphalipoproteinemia, hypercholesterolemia, hypolipemia, atherosclerosis, diabetes and nephrotic syndrome. For instance, it is demonstrated experimentally that several times higher levels of CETP are secreted in blood of the patients with hyperlipemia compared to that of the healthy volunteers. On the other hand, CETP, HDL-cholesterol and atherosclerosis have the following relationship. When low CETP activity hardly causes atherosclerosis, but with high level of HDL cholesterol. In contrast, when high CETP activity causes atherosclerosis and with low level of HDL cholesterol. Such relationship is experimentally demonstrated(Current Therapy, vol. 7, 9:36-45(1989)). In order to elucidate the relationship, assay methods for CETP in body fluid such as blood plasma from healthy person or patient having the above various diseases, especially immunoassays such as radio immunoassay(RIA) by using monoclonal antibody against CETP(anti-CETP monoclonal antibody), or enzyme immunoassay(EIA, ELISA) is being developed together with anti-CETP monoclonal antibody for the assay methods. As to EIA(ELISA) assay with the anti-CETP monoclonal antibody, for examples, Imai et al.(Japanese Unexamined Patent publication NO.HEI6-169793), Nakano, et al.,(Arteriosclerosis, vol. 19, 11:951, No. 22, (1991)), Takahama, et al.(Arteriosclerosis, vol. 20, 10:837, No. 135, (1992)), Sato, et al.(Arteriosclerosis, vol. 20, 10:836, No. 134, (1992)), H. Mezdur, et al.(Clinical Chemistry, vol. 40, 4:593-597(1994)) Clark et al.(Journal of Lipid Research, vol. 36,:876(1995)) reported sandwich ELISA method by using two kinds of anti-CETP monoclonal antibodies or their Fab". However, these assay methods requires complicated handling. Before assay, plasma sample has to be heat-treated at 95-100°C and/or pre-treated with a detergent such as Tween- 20 and Triton X-100. As heat treatment, of plasma sample causes denaturation of CETP in the sample, the assay result tells only the amount of denatured CETP. So, it is impossible to assay intact CETP in plasma sample accurately. Also, R. Clark et al.(FASEB Journal, vol. 8, 7:A1343 No. 495(1994)), Takahashi et al.(Arteriosclerosis, vol. 20, 10:837 No. 136(1992) and ibid. vol. 21, 3:209 No. 97(1993)), Kanamitu et al.(ibid. vol. 21, 3:209 No. 98(1993)), Waki et al.(ibid. vol. 22, 5:441 No. 194(1994)) reported the assay results on CETP in plasma samples by ELISA, however, they did not described in detail preparation methods and properties of anti-CETP antibodies and specific assay procedures. Regarding RIA assay methods with the anti-CETP monoclonal antibody, Fukazawa et al.(Study on lipid biochemistry, vol. 34:163-166(1992)), Y. Marcel et al.(J. Clin. Invest., vol. 85:10-17(1990) and Adv. Exp. Med. Biol., vol. 243:225- 230(1988)), Fukazawa et al.(J. Biochem., vol. 111:696- 698(1992)), J.Koizumi et al.(Atherosclerosis, 90:189- 196(1991), M. Brown et al. (Nature, vol. 342:448-451(1989)-) and V. Dangremont et al.(Clinica Chimica Acta, vol. 231:147- 160(1994)) published reports. However, these RIA methods also had defects as EIA(ELISA) methods had. The defects were complexity of handling (the methods required pre-treatment of plasma sample with a detergent) and/or insufficient sensitivity for detection. As described above, many researchers tried to establish assay method for CETP- However, there is no report that such a convenient and sensitive system which needs no pretreatment of a sample and enables to determine conveniently the amount of intact CETP was established. To elucidate relationship between the diseases (such as atherosclerosis, hyperlipemia and hyper high density lipoproteinemia) and CETP that plays important role in the cholesterol reverse transfer system which may be highly related to the onset of the diseases, it is strongly requested to develop a simple and sensitive assay method that can be widely applicable to clinical purposes to assay intact CETP in body fluid including blood plasma from healthy person or any patient, and to develop monoclonal antibody used for said assay method. However, they have not been established yet. The present invention provides an assay method widely applicable for clinical purposes and the anti-human CETP monoclonal antibody which is very useful not only for said assay method but also as a reagent for separation/purification of CETP and for a pharmaceutical composition. SUMMARY OF THE INVENTION The inventors of the present invention investigated , thoroughly on assay methods for CETP in human body fluid that can be widely applicable for clinical purposes and on monoclonal antibodies against human CETP used for said assay methods, and, by using biologically active purified human CETP as an inununogen were succeeded in preparing three anti-human CETP monoclonal antibodies which respectively has different high binding specificity(CETP inhibitory activity) to human CETP, especially intact CETP in human body fluid. Furthermore, as said three anti-human CETP monoclonal antibodies have high binding specificity(CETP inhibitory activity) to human CETP, especially intact CETP in human body fluid, the inventors found that, for assaying intact CETP in human body fluid(blood plasma etc.)/ the immunoassay using the anti-human CETP monoclonal antibodies of the present invention can provide a simple and sensitive assay method which have not been established so far, and thus the inventors completed the present invention. Particularly, as the three anti-human CETP monoclonal antibodies have high but different binding specificity(CETP inhibitory activity) to human CETP(especially to intact CETP in human body fluid), by using combination of any of the two of the said three monoclonal antibodies for sandwich ELISA, the inventors were succeeded in developing simpler and more sensitive assay method which can be widely applicable for clinical purposes. As the anti-human CETP monoclonal antibodies have high binding specificity(CETP inhibitory activity) to human CETP(especially to intact CETP in human body fluid), by using the monoclonal antibodies of the invention, it is possible to assay human CETP simply and sensitively without any pre- treatment such as heat treatment or detergent treatment of a plasma sample. In addition to that, as the anti-human CETP monoclonal antibodies of the invention have high CETP inhibitory activity, they are useful as Pharmaceuticals for treatment or prevention of various diseases caused by abnormal dynamics such as atherosclerosis, hyperlipemia, high blood HDL and hypercholesterolemia. The first aspect of the present invention is a monoclonal antibody having at least the following characteristics of: (a) Reactive to plasma CETP(Cholesterol Ester Transfer Protein) of healthy human, hyperlipemia patients, LCAT(Lecithin Cholesterol Acyl Transferase) deficiency patients or hyper HDL patients. (b) Not reactive to rabbit CETP at the concentration of 3 µg/ml or below. (c) Not specially reactive to denatured human CETP. The second aspect of the present invention is the hybridomas which produce monoclonal antibodies reactive to human CETP, more specifically, they are hybridomas #72-1 and #86-2 identified by the Accession Number FERM BP-4944 and FERM BP-4945 respectively. The third aspect of the present invention is the monoclonal antibodies reactive to human CETP, more specifically, they are monoclonal antibodies #72-1 and #86-2, and are derived respectively from the hybridomas #72-1 and #86-2 identified by the Accession Number FERM BP-4944 and FERM BP-4945 respectively. The fourth aspect of the present invention is recombinant chimeric monoclonal antibody reactive to human CETP comprising a variable region derived from that of the above mentioned monoclonal antibody and a constant region derived from that of human immunoglobulin. The fifth aspect of the present invention is the recombinant humanized monoclonal antibody reactive to human CETP, in which a part of or the whole of the complementarity determining regions of the hypervariable region are derived from that of the above mentioned monoclonal antibody, the framework regions of the hypervariable region are derived from that of human immunoglobulin, and the constant region is derived from that of human immunoglobulin. The sixth aspect of the present invention is the antibody fragment termed F (ab") 2 or Fab" derived from the above mentioned monoclonal antibody, the above mentioned recombinant chimeric monoclonal antibody or the above mentioned recombinant humanized monoclonal antibody. The seventh aspect of the present invention is the immobilized monoclonal antibody and the immobilized antibody fragment. The immobilized monoclonal antibody is prepared by immobilizing the monoclonal antibody, the recombinant chimeric monoclonal antibody or the recombinant humanized monoclonal antibody on an insoluble carrier. The immobilized antibody fragment is prepared by immobilizing the above mentioned antibody fragment F(ab")2 or Fab" on an insoluble carrier. More specifically, the immobilized monoclonal antibody or the immobilized antibody fragment immobilized on a plate, test tube, tube, beads, ball, filter, membrane or insoluble carrier used for affinity column chromatography. The eighth aspect of the present invention is the labeled monoclonal antibody and the labeled antibody fragment. The labeled monoclonal antibody is prepared by labeling the monoclonal antibody, the recombinant chimeric monoclonal antibody or the recombinant humanized monoclonal antibody with a substance capable of providing detectable signal independently or by reaction with other substance. The labeled antibody fragment is prepared by labeling the F(ab")2 or Fab" with a substance capable of providing detectable signal independently or by reaction with other substance. More specifically, they are the labeled monoclonal antibody or the labeled antibody fragment labeled with an enzyme, a fluorescent material, a chemical luminous material, biotin, avidin or radioisotopes. The ninth aspect of the present invention is a kit for assay or detection of human CETP comprising at least the monoclonal antibody, the recombinant. chimeric monoclonal antibody or recombinant humanized monoclonal antibody, the above mentioned antibody fragment F(ab")2 or Fab", the immobilized monoclonal antibody or immobilized antibody fragment, or the labeled monoclonal antibody or the labeled antibody fragment. Specifically, a kit for assay or detection of human CETP containing the immobilized monoclonal antibody or the immobilized antibody fragment and the labeled monoclonal antibody or the labeled antibody fragment. The tenth aspect of the present invention is a method for assay and detection of human CETP by immunoassay characterized by using at least of the monoclonal antibody, the recombinant chimeric monoclonal antibody or the recombinant humanized monoclonal antibody, the antibody fragment F(ab")2 or Fab", the immobilized monoclonal antibody or the immobilized antibody fragment, or the labeled monoclonal antibody or the labeled antibody fragment. BRIEF DESCRIPTION OF THE ACCOMPANYING DRAWINGS Fig. 1 shows the chromatogram of Phenyl Sepharose column chromatography of human CETP. Fig. 2 shows the chromatogram of Resource Q column chromatography of human CETP. Fig. 3 shows the chromatogram of Affinity column chromatography by using the monoclonal antibody #72-1 of human CETP. Fig. 4 shows the molecular weight of human CETP purified the affinity column chromatography by using the oclonal antibody #72-1. Fig. 5 shows the reactivity of the monoclonal antibodies, #72-1, #86-2, and #176-1 for the purified human CETP. Fig. 6 shows the chromatogram by Phenyl Sepharose column chromatography of rabbit CETP. Fig. 7 shows the chromatogram by Blue Sepharose column chromatography of rabbit CETP. Fig. 8 shows the chromatogram by the succinylated LDL column chromatography of rabbit CETP. Fig. 9 shows the reactivity of the monoclonal antibodies, #72-1, #86-2, and #176-1 for the purified rabbit CETP. Fig. 10 shows the sensitivity of the determination when the labeled monoclonal antibody #86-2 and several immobilized antibodies are combined. Fig. 11 shows the sensitivity of the determination when the labeled monoclonal antibody #176-1 and several immobilized antibodies are combined. Fig. 12 shows the sensitivity of the determination when the labeled monoclonal antibody #72-1 and several immobilized antibodies are combined. Fig. 13 shows the sensitivity of the determination when the labeled monoclonal antibody #86-2 and several concentration of the immobilized antibody #72-1 are combined. Fig. 14 shows the sensitivity of the determination when the labeled monoclonal antibody #72-1 and several concentration of the immobilized antibody #86-2 are combined. Fig. 15 shows the sensitivity of the determination when the labeled monoclonal antibody #176-1 and several concentration of the immobilized antibody #72-1 are combined. Fig. 16 shows the sensitivity of the determination when the labeled monoclonal antibody #72-1 and several concentration of the immobilized antibody #176-1 are combined. Fig. 17 shows the sensitivity of the determination when the labeled monoclonal antibody #72-1(1 µg/ml) and the immobilized antibody #86-2(1 ng/well or 3 ng/well) are combined. Fig. 18 shows a calibration curve of the purified human CETP standard preparations obtained from the determination method of the present invention. Fig. 19 shows a calibration curve of the human CETP standard preparations from human plasma with different purity obtained from the determination method of the present invention. Fig. 20 shows a correlation between the amount determined and the activity of CETP in the samples by using the determination method of the present invention. Fig. 21 shows the amount determined and the activity of plasma CETP from normal subjects or various patients by using the determination method of the present invention. Fig. 22 shows the reactivity of the monoclonal antibodies, #72-1 or #86-2 for the denatured human CETP, and the difference of the determination method between the present invention and conventional one. Fig. 23 shows the CETP inhibition effect of anti-CETP monoclonal antibody in vivo test of rabbit. Fig. 24 shows the CETP inhibition effect of anti-human CETP monoclonal antibody in vivo test of transgenic mice which highly express human CETP. Fig. 25 shows the effect to the HDL choresterol level in plasma from anti-human CETP antibody in vivo test. DETAIL DESCRIPTION OF THE INVENTION The first specific aspect is a method for assay or detection of human CETP by immunoassay comprising at least the following steps of: (a)reacting a sample with the above mentioned immobilized monoclonal antibody or the immobilized antibody fragment; and (b)reacting the labeled monoclonal antibody or the labelOed antibody fragment with antigen-antibody complex. The antigen-antibody complex formed by binding human CETP in a sample with said immobilized monoclonal antibody or said immobilized antibody fragment. The second specific aspect is a method for assay or detection of human CETP by immunoassay comprising at least the following steps of: (a) reacting a sample with the above mentioned labeled monoclonal antibody or the labeled antibody fragment; and (fc>) reacting the above mentioned immobilized monoclonal antibody or the immobilized antibody fragment with antigen- antibody complex. The antigen-antibody complex formed by binding human CETP in a sample with said labeled monoclonal antibody or said labeled antibody fragment. The third specific aspect is a method for assay or detection of human CETP by immunoassay comprising at least the following step of: (a) reacting a mixture comprising the immobilized monoclonal antibody or the immobilized antibody fragment, the labeled monoclonal antibody or the labeled antibody fragment and a sample. The fourth specific aspect is a method for assay or detection of human CETP by immunoassay comprising at least the following step of: (a) reacting a sample and the labeled human CETP standard labeled with a labeling substance capable of providing detectable signal independently or by reaction with other substance with the above mentioned immobilized monoclonal antibody or the immobilized antibody fragment. The fifth specific aspect is a method for assay or detection of human CETP by immunoassay comprising at least the following step of (a) or following steps of (b) and (c): (a) reacting the above mentioned monoclonal antibody or antibody fragment with a mixture of a sample and the labeled human CETP standard. The labeled human CETP standard is prepared by labeling with a substance capable of providing detectable signal independently or by reaction with other substance; (b) reacting the above mentioned monoclonal antibody or the antibody fragment with a sample; (c) followed by the step (b), reacting the labeled human CETP standard with the reaction mixture of step (b). The labeled human CETP standard is prepared by labeled with a substance capable of providing detectable signal independently or by reaction with other substance. More specifically, a method for assay or detection of human CETP by immunoassay comprising at least the following steps of (a) and (d), or steps of (b) to (d): (a) reacting the monoclonal antibody or the antibody fragment of the present invention with a mixture of a sample and the labeled human CETP standard. The labeled human CETP standard is prepared by labeling with a substance capable of providing detectable signal independently or by reaction with other substance; (b) reacting the monoclonal antibody or the antibody fragment of the present invention with a sample; (c) followed by the step (b), reacting the labeled human CETP standard with the reaction mixture of step (b). The labeled human CETP standard is prepared by labeling with a substance capable of providing detectable signal independently or by reaction with other substance; (d) reacting an anti-serum derived from mammals reactive to said monoclonal antibody or said antibody fragment with an antigen-antibody complex. The antigen-antibody complex is formed by binding human CETP in said sample or said labeled human CETP standard with said monoclonal antibody or said antibody fragment. The eleventh aspect of the present invention is a kit for separation or purification of human CETP comprising the immobilized monoclonal antibody or the immobilized antibody fragment. The twelfth aspect of the present invention is a method for separation or purification of human CETP by affinity chromatography by using the immobilized monoclonal antibody or the immobilized antibody fragment. More specifically, a purification method for human CETP by using column chromatography. The thirteenth aspect of the present invention is a mouse introduced DNA encoding a human CETP which constantly secretes human CETP in blood without extrinsic or artificial induction. The fourteenth aspect of the present invention is a pharmaceutical composition comprising any one of the monoclonal antibody, the recombinant chimeric monoclonal antibody, the recombinant humanized monoclonal antibody or the antibody fragment (F(ab")2 or Fab"), and a pharmaceutically acceptable carrier. More specifically, the pharmaceutical composition for treating and/or preventing hyperlipemia or atherosclerosis. By clarifying the meanings of the terms used here, the present invention is explained in detail. "A monoclonal antibody" of the present invention means a monoclonal antibody reactive to human CETP and more specifically, monoclonal antibody having at least the following characteristics (a), (b) and (c). (a) Reactive to plasma CETP (Cholesterol Ester Transfer Protein) of healthy human, hyperlipemia patients, LCAT(Lecithin Cholesterol Acyl Transferase) deficiency patients or hyper HDL(high density lipoprotein) patients. (b) Not reactive to rabbit CETP at the concentration of 3 µg/ml or below. (c) Not specifically reactive to denatured human CETP. In addition, the monoclonal antibody of the present invention has a characteristic that it is reactive to intact CETP in human body fluid. "Denatured human CETP" mentioned here means human CETP of which higher-order structure(conformation) of protein has destroyed due to such as heat treatment or treatment with detergent(Tween-20, Triton X-100 etc.), or which is biologically inactive. More specifically, "monoclonal antibody" means the monoclonal antibodies #72-1 and #86-2 derived from the hybridomas #72-1 and #86-2 identified by Accession Number FERM BP-4944 and FERM BP-4945 respectively, and the monoclonal antibody #176-1 produced from the hybridoma #17 6-1 prepared in the example described below. Furthermore, it includes a recombinant chimeric monoclonal antibody, a recombinant humanized monoclonal antibody which can be prepared by genetic recombination, derived from said monoclonal antibody, and also includes human monoclonal antibody. The monoclonal antibodies of the present invention(for example, #72-1, #86-2 and #176-1) can be prepared by conventional preparation methods established for monoclonal antibodies. For instance, they can be produced from hybridomas prepared by cell fusion. That is, by immunizing mammals with antigen, antibody producing cells can be obtained. Hybridoma cells are prepared with said antibody producing cells and myeloma cells which does not have ability to produce autoantibody. After cloning of said hybridoma, a clone which produces monoclonal antibody showing specific affinity to the antigen used for immunization of the mammals is selected and then, the monoclonal antibody of the present invention is produced from the hybridoma. Specifically, non-denatured, intact, and biologically active purified human CETP is used as an antigen. Mammal (including transgenic animals such as genetically engineered mouse which produces human antibody), more specifically, mouse, rat, hamster or guinea pig, is immunized with said antigen by injecting said antigen once to several times subcuteneously, intramuscularly, intravenously, into food pad or intraperitoneally. In general, immunization is performed once to four times every one to fourteen days from the initial immunization. Antibody producing cells are obtained from said immunized mammal approximately one to five days after the final immunization. The hybridoma which secretes monoclonal antibody can be prepared according to the method by Kohler, and Milstein et al.(Nature, vol. 256:495-497(1975)) or according to modified method similar to the original method. Hybridoma is prepared by cell fusion of antibody producing cells and myeloma cells. The antibody producing cells are contained in spleen, lymph node, bone marrow or tonsil, preferably in spleen, which is obtained from the mammals immunized according to the above mentioned procedures. The myeloma cells incapable of producing autoantibody are derived from mammals preferably from mouse, rat, guinea pig, hamster, rabbit or human, more preferably from mouse, rat or human. As myeloma cells to be used for cell fusion, for example, mouse derived myeloma P3/X63-AG8..653(653), P3/NSl/l-Ag4-l(NS- 1), P3/X63-Ag8.Ul(P3Ul), SP2/O-Agl4(Sp2/O, Sp2), PAI, FO or BW5147, rat derived myeloma 210RCY3-Ag.2.3., human derived myeloma U-266AR1, GM1500-6TG-A1-2, UC729-6, CEM-AGR, D1R11 or CEM-T15 and so forth can be used. Screening of hybridoma can be performed by culturing the hybridoma on microtiter plate as an example, then, by measuring reactivity of the culture supernatant of the well which shows proliferation to purified human CETP used for the immunization of mammals by ria or by enzyme immunoassay such as ELISA. Preparation of monoclonal antibody from hybridoma is performed by culturing the hybridoma in vitro or by placing the hybridoma in vivo in ascites of mouse, rat, guinea pig, hamster or rabbit, preferably in ascites of mouse or rat, more preferably in ascites of mouse, and then, by isolating monoclonal antibody from the culture supernatant or from the ascites of mammals. In case of culturing hybridoma in vitro, depending on the characteristics of cells, purpose of study, culturing method and so forth, known nutrient media or any kind of nutrient media modified from known basic media can be used as long as the media is usable to proliferate, maintain and keep the hybridoma and to produce monoclonal antibody in culture supernatant. As a basic media, for example, low calcium media such as Ham"Fl2 media, MCDB153 media and low calcium MEM media and high calcium media such as MCDB104 media, MEM media, D-MEM media, RPMI1640 media, ASF104 media or RD media can be applied. Depending on purpose of culture, said basic media may contain, for example, serum, hormones, cytokines and/or various inorganic or organic substances. Isolation and purification of the monoclonal antibody from the above mentioned culture supernatant or ascites can be performed by applying to saturated ammonium sulfate precipitation, euglobulin precipitation, caproic acid method, caprylic acid method, ion exchange chromatography(DEAE, DE52 or the like), affinity column chromatography such as anti- immunoglobulin column chromatography and protein A column chromatography, and so forth. "Recombinant chimeric monoclonal antibody" of the present invention means genetically engineered monoclonal antibody, specifically chimeric monoclonal antibody such as recombinant mouse/human chimeric monoclonal antibody characterized by its variable region derived from that of the above mentioned monoclonal antibody such as the monoclonal antibody #72-1, #86-2 or #176-1 and its constant region derived from that of human immunoglobulin. The constant regions derived from human immunoglobulins have different proper amino acid sequences based on their isotype such as IgG, IgM, IgA, IgD and IgE. The constant region of human immunoglobulin of any isotype can be applicable to the constant region of the recombinant chimeric monoclonal antibody of the present invention. The constant region of IgG is preferable. Recombinant mouse/human chimeric monoclonal antibody derived from the monoclonal antibody #72-1, #86-2 or #176-1 included in the monoclonal antibodies of the present invention can be prepared as follows, however, it is not limited to the following preparation method. The recombinant chimeric monoclonal antibody can be prepared by, for example, referring Experimental Medicine (special edition) vol. 1-6, No. 10, 1988 and Japanese patent publication No. HEI 3-73280. At the downstream of the active VH gene(rearranged VDJ gene encoding variable region of H chain), the CH gene(C gene encoding constant region of H chain) is operably linked to, and at the downstream of the active VL gene(rearranged VJ gene encoding variable region of L chain), the CL gene(C gene encoding constant region of L chain) is operably linked to. These genes are inserted in same or different expression vector. The vh gene used here is obtained from DNA encoding the monoclonal antibody #72-1, #86- 2 or #176-1 isolated from the hybridoma #72-1, #86-2 or #176-1 of the present invention. The CH gene and the CL gene are obtained from DNA encoding human immunoglobulin. The VL gene is obtained from DNA encoding the monoclonal antibody #72-1, #86-2 or #176-1 isolated from the hybridoma #72-1, #86-2 or #176-1 of the present invention. Host cells are transformed with said vector(s). The recombinant mouse/human chimeric monoclonal antibody of the present invention can be prepared by culturing said transformed cells. As a host cell, prokaryotic cells (e.g. E. coli) or eukaryotic cells (e.g. CHO cell) can be used. Briefly, after DNA is extracted from the hybridomas #72- 1, #86-2 or #176-1 by a conventional method, said DNA is digested with an appropriate restriction enzyme such as EcoRI and ffindlll. Then, southern blotting is performed by electro- phoresis (for example, with 0.7% agarose gel). After the electrophoresis, the gel is stained(for example, with etidium bromide). After taking photograph of the gel, location of marker is marked and the gel is washed twice, then, the gel is immersed in 0.25 M HC1 solution for 15 minutes. Then the gel is immersed in 0.4 N NaOH for 10 minutes with gentle shaking. By a conventional method, the gel is transferred on a filter and after 4 hours standing, the filter is recovered and is washed twice with 2 x SSC. After drying the filter completely, baking filter is performed at 75°C for 3 hours. After the baking, said filter is put in 0.1 x SSC/0.1 % SDS solution and is treated for 30 minutes at 65°C. The filter is immersed in 3 x SSC/0.1 % SDS solution. Then, the filter is put in a vinyl bag with the pre-hybridization solution and stand for 3 to 4 hours at 65°C. Then, 32p labeled probe DNA and the hybridization solution are added, and the mixture is reacted for approximately 12 hours at 65°C. After hybridization, the filter is washed at appropriate salt concentration, reaction temperature and time period(as an example, with 2 x SSC-0.1 % SDS solution, at room temperature for 10 minutes). Said filter is then put in a vinyl bag and small amount of 2 x SSC is added, the bag is closed tightly and autoradiography is performed. By the above-mentioned southern blotting, the rearranged VDJ gene and VJ gene which respectively encode H chain and L chain of the monoclonal antibody #72-1, #86-2 or #176-1 are identified. The region including the identified DNA fragment is fractionated by sucrose density gradient centrifugation and incorporated into phage vector(such as Charon 4A, Charon 28, X.EMBL3 and XEMBL4). By transforming E. coli(such as LE392 and NM539) with said phage vector, genome library is established. By plaque hybridization with an appropriate probe(such as H chain J gene and L chain(k) J gene) according to Benton-Davis method(Science, vol. 196, p.180-182, 1977), positive clones containing rearranged VDJ gene or VJ gene respectively are obtained from the genome library. On the obtained clones, the restriction enzyme map is made, the nucleotide sequence is determined and existence of the rearranged VH(VDJ) gene or VL(VJ) gene is confirmed. On the other hand, human CH gene and human CL gene used for chimerization are isolated separately. For instance, to prepare chimeric antibody with human IgGl, C?l gene(CH gene) and Ck gene(CL gene) are isolated. These genes can be isolated from human genome library by using mouse C?l gene(corresponds to human C?l gene) and mouse Ck gene(corresponds to human Ck gene) as probes because the nucleotide sequences of mouse immunoglobulin gene and human immunoglobulin gene are highly homologous. Briefly, as an example, a DNA fragment containing human Ck gene and the enhancer region can be isolated from HaeIII- Alul genome library of the XCharon 4A (Cell, vol. 15:1157- 1174(1978)) by using HindIII-BamHI fragment(3 kb) obtained from the clone Igl46(Proc. Natl. Acad. Sci. USA, vol. 75:4709- 4713(1978) and EcoRI fragment(6.8 kb) obtained from the clone MEPlO(Proc. Natl. Acad. Sci. USA, vol. 78:474-478(1981)) as probes. The human C?l gene can be isolated by, for example, digesting DNA from human fetal liver cell with Hindlll, fractionating by agarose gel electrophoresis, inserting 5.9 kb band into X788 and then isolating with the above mentioned probes. Using the isolated a mouse VH gene, mouse VL gene, human CH gene and human CL gene, and considering the promoter region and the enhancer region, human CH gene is placed at the downstream of mouse VH gene and human CL gene is placed at the downstream of mouse VL gene in a expression vector such as pSV2gpt and pSV2neo by using an appropriate restriction enzyme and DNA ligase. In this case, mouse VH gene/human CH gene and mouse VL gene/human CL gene can be placed in one expression vector or in different expression vectors. The expression vector(s) inserted chimeric gene is introduced in myeloma cells which do not produce antibody such as P3X63-Ag8-653 cell and SP210 cell by protoplast fusion method, DEAE-dextran method, calcium phosphate method or electrical punching method. The transformed cells are selected by culturing them in culture media containing a drug which corresponds to the drug resistant gene inserted in the expression vector(s), and chimeric monoclonal antibody producing cells can be obtained. From the culture supernatant of the selected antibody producing cells, the chimeric monoclonal antibody can be obtained. "A recombinant humanized monoclonal antibody" of the present invention means genetically engineered monoclonal antibody, specifically means that a part of or the whole of the complementarity determining regions of hypervariable region is derived from that of the above mentioned monoclonal antibody(monoclonal antibody #72-1, #86-2 or #176-1 as examples), the framework regions hypervariable region are derived from that of human immunoglobulin, and the constant region is derived from that of human immunoglobulin. The complementarity determining region(s) in the hypervariable region means a part of or the whole of the three regions(CDR1, CDR2, CDR3) which exist in the hypervariable region of the variable region of antibody and complementarily binds directly with antigen. The framework region(s) in super variable region means the four regions(FR1, FR2, FR3, FR4) which are located before and after said three complementarity determining regions, and are relatively well maintained (constant). In other words, "Recombinant humanized monoclonal antibody" means monoclonal antibody in which all of the parts except for a part of or the whole of the complementarity determining regions of the hypervariable region of the monoclonal antibodies of the present invention are replaced with corresponding region(s) of human immunoglobulin. The constant region derived from human immunoglobulin has its proper amino acid sequence depending on its isotype such as IgG, IgM, IgA, IgD and IgE. The constant region of the recombinant humanized monoclonal antibody of the present invention may be the constant region of any isotype of human immunoglobulin. Preferably, the constant region of human IgG. And also, the framework region(s) in the hypervariable region derived from human immunoglobulin is not limited to one isotype. The recombinant humanized monoclonal antibody derived from the monoclonal antibody #72-1, #86-2 or #176-1 included in the monoclonal antibodies of the present invention can be prepared by, for example, the following steps but not limited to the following. For instance, that can be prepared by genetic engineering technique according to Japanese Patent Unexamined Publication Number HEI4-506458 and Japanese Patent Unexamined Publication Number SYOU62-296890. From the hybridoma #72-1, #86-2 or #176-1, at least one mouse H chain CDR gene and at least one mouse L chain CDR gene which corresponds to said mouse H chain CDR gene are isolated. From human immunoglobulin gene, human H chain gene which encodes whole region except human H chain CDR(corresponds to mouse H chain CDR), and human L chain gene which encodes whole region except human L chain CDR(corresponds to mouse L chain CDR) are isolated. Said isolated mouse H chain CDR gene and said H chain gene are operably linked and are introduced into an appropriate expression vector, and said mouse L chain CDR gene and said human L chain gene are operably linked and are introduced into another appropriate expression vector. Or otherwise, it is possible to introduce said mouse H chain CDR gene/human H chain gene and mouse L chain CDR gene/human L chain gene into the same one expression vector. By transforming host cells with the obtained expression vector(s), transformed cells which produce humanized monoclonal antibody can be obtained. By culturing said transformed cells, human monoclonal antibody can be obtained from culture supernatant. As host cells, both prokaryotic cells such as E. coll and eukaryotic cells such as CHO(Chinese hamster ovarian) cells may be used. The term "human antibody" in the present invention means that the immunoglobulin comprises entire regions including both constant and variable regions of heavy chains and those of light chains, and that it is derived from genes encoding human immunoglobulin. The human antibody is produced according to the common method. In general, the human immunoglobulin genes are incorporated into gene locus of the animals such as mice except human to obtain transgenic animals producing humanized antibody. Then, the transgenic animal is immunized by using any antigen. Thus, polyclonal and/or monoclonal antibodies are obtained. For instance, such transgenic mice producing human antibodies are obtained by using the method disclosed in several references: Nature Genetics, 7:13-21(1994), NIKKEI Science, June: 40-50(1995), Nature, 368:856-859(1994), and Japanese National Publication of translation for the foreign language patent application No. 1995-500233. "F(ab")" or "Fab1" of the present invention mean antibody fragments produced by treating immunoglobulin (monoclonal antibody) with proteinase such as pepsin and papain. They mean the antibody fragment(s) digested at the positions before and after the disulfide bonds between the two H chains in the hinge region. For example, when IgGl is treated with papain, it is cleaved at the upstream of the disulfide bond between the two H chains in the hinge region to make two identical fragments, i.e., L chain consisting of VL(L chain variable region) and CL(L chain constant region), and H chain fragment consisting of VH(H chain variable region) and CH?1(?1 region of the constant region of H chain) are bound at the C terminal region by disulfide bond. Each of the two identical fragments is called Fab". When treated with pepsin, IgGl is cleaved at the downstream of the disulfide bond between the two H chains at the hinge region to make one antibody fragment which is a little larger than a fragment of two Fab" connected at the hinge region. This antibody fragment is called F(ab")2- "Anti serum derived from mammal" of the present invention means serum containing antibody reactive to the monoclonal antibody or the its antibody fragment of the present invention. The serum is prepared by immunizing mouse, rat, guinea pig, rabbit, goat, pig or cattle, preferably rat, guinea pig, rabbit or goat, with said monoclonal antibody or said antibody fragment of the present invention according to the procedures described above for preparation of the above mentioned monoclonal antibody. Specifically, antiserum derived from said mammals immunized with the above mentioned monoclonal antibody(for example monoclonal antibody #72-1, #86-2 or #176-1) or antibody fragment F(ab")2 or Fab" of these antibodies. "Insoluble carrier" of the present invention means a carrier which is used in order to carry human CETP in a sample(for example, body fluid sample such as blood plasma, culture supernatant or centrifuge supernatant) or the above mentioned monoclonal antibody or the antibody fragment by, for example, physical adsorption or chemical bond. As examples, the following can be used; (1) plastics composed of polystyrene resin, polycarbonate resin; silicon resin or nylon resin; plate composed of water insoluble substances such as glass; test tube or tube having inner capacity; beads, ball, filter or membrane etc., and (2) insoluble carriers used for affinity chromatography such as cellulose type carrier, agarose type carrier, polyacrylami.de type carrier, dextran type carrier, polystyrene type carrier, polyvinyl alcohol type carrier, poly amino acid type carrier and porous silica carrier. "Immobilized monoclonal antibody" or "immobilized antibody fragment" of the present invention means the monoclonal antibody or the antibody fragment which is bound to said insoluble carrier by physical adsorption or chemical bond. These immobilized monoclonal antibody or the immobilized antibody fragment can be used for detection, assay, separation and purification of human CETP in sample(for example, body fluid sample such as blood plasma, culture supernatant and centrifugation supernatant). For the purpose of said detection or assay, the immobilized monoclonal antibody or immobilized antibody fragment immobilized on the insoluble carrier mentioned above (1). Especially for assay, considering simple handling and simultaneous handling of many samples, it is preferable to use a plastic plate having many wells such as microtiter plate with 96 wells. For said separation and purification, the immobilized monoclonal antibody or immobilized antibody fragment immobilized on a filter mentioned above (1) or a membrane mentioned above (2) can be used. "Labeling substance capable of providing detectable signal independently or by reaction with other substance" of the present invention mean a substance which is used to detect the above mentioned monoclonal antibody, the antibody fragment or the human CETP standard. They are detectable by binding said substance with the antibody, the fragment or the standard (physicochemical binding etc.), Specifically, the substance is enzyme, fluorescent material, chemical luminous material, biotin, avidin or radioisotope, and so forth. More specifically, the substance is enzyme such as peroxidase, alkaline phosphatase, ß-D-galactosidase, glucose oxidase, glucose-6-phosphate dehydrogenase, alcohol dehydrogenase, malate dehydrogenase, penicillinase, catalase, apo-glucose oxidase, urease, luciferase and acetylcholine esterase; fluorescent material such as fluorescence isothiocyanate, phycobilin protein, rare earth metal chelate, dansylchloride and tetramethylrhodamin isothiocyanate; radioisotope such as 3H, 14C and 131I. biotin, avidin or chemical luminous materials. Radioisotopes and fluorescent materials can provide detectable signal independently. On the other hand, enzymes, chemical luminous materials, biotin and avidin can not provide detectable signal independently, and with one or more other substance(s), they can provide detectable signal. For instance, in the case of enzyme, it requires at least its substrate. Various substrates are used depending on method for measuring enzyme activity(Colorimetric method, fluorescence method, bio-luminescence method or chemical luminescence method etc.). In the case of biotin, generally, it is reacted with, at least, avidin or enzymatically modified avidin, but not limited to the method. If it necessary, various coloring developing substance depending on said substrates are used. "Labeled monoclonal antibody", "labeled antibody fragment" and "labeled human CETP standard" of the present invention mean the monoclonal antibody, the antibody fragment and the human CETP standard labeled with said labeling substance, respectively. These labeled monoclonal antibody, labeled antibody fragment and labeled human CETP standard can be used for detection and assay of human CETP in a sample such as body fluid sample(blood plasma etc.), culture supernatant and centrifugation supernatant. In the present invention, any labeling substance mentioned above can be used, however, considering detection limit(sensitivity), assay limit and handling(convenience), biotin labeling is preferable. "Human CETP standard" of the present invention means human CETP used as a standard for detection or assay of human CETP in a sample as mentioned above. "Immunoassay" of the present invention means a method for detection or assay of an antigen contained in a sample(for example, body fluid sample such as blood plasma, culture supernatant and centrifugation supernatant) based on the principle of antigen-antibody reaction. In the present invention, any known immunoassay methods can be applicable to the present invention as long as antibody used for said antigen-antibody reaction is one or more said monoclonal antibody or antibody fragment selected from the above mentioned monoclonal antibody or antibody fragment, the above mentioned immobilized monoclonal antibody or the immobilized antibody fragment the above mentioned labeled monoclonal antibody or the labeled antibody fragment, and as antigen assayed is human CETP. Specifically, the methods described in "Enzyme Immuno Assay"(3rd edition, Eiji Ishikawa et al. ed., Igakusyoin, 1987) such as single antibody solid phase method, double antibody liquid phase method, double antibody solid phase method, sandwich method, enzyme multiplied immunoassay technique(EMIT), enzyme channeling immunoassay, Enzyme modulator mediated enzyme immunoassay (EMMIA), enzyme inhibitor immunoassay, immunoenzymometric assay, enzyme enhanced immunoassay and proximal linkage immunoassay, or one pot method as described in Japanese Patent Publication No. HEI2-39747. In the present invention, depending on purpose of assay, appropriate immunoassay can be selected. Considering easily handling and/or economical efficiency, especially considering wide clinical use, sandwich method, one pot method, single antibody solid phase method or double antibody liquid phase method is preferable. It is more preferable to use sandwich method or one pot method. It is most preferable to use sandwich method with the immobilized monoclonal antibody or the immobilized antibody fragment immobilized on a microplate having many wells such as 96 well microplate and the labeled monoclonal antibody or the labeled antibody fragment labeled with enzyme or biotin, or otherwise, one pot method with the immobilized monoclonal antibody or the immobilized antibody fragment immobilized on beads or ball and labeled monoclonal antibody or labeled antibody fragment labeled with enzyme or biotin. A specific example of the most preferable embodiment is a sandwich method or one pot method using the following combination, the monoclonal antibody #72-1, #86-2 or #176-1, or their F(ab")2 or Fab1 immobilized on a microplate, beads or ball and with the monoclonal antibody #72-1, #86-2 or #176-1 or their F(ab")2 or Fab1 labeled with enzyme or biotin. Although the monoclonal antibodies #72-1, #86-2 and #176-1 can be used for immobilized form as well as labeled form, it is preferable to use #72-1 as immobilized form and #86-2 or #176- 1 as labeled form. Particularly, it is more preferable to use #72-1 (which is the monoclonal antibody derived from the hybridoma identified with FERM BP4944) as immobilized monoclonal antibody and #86-2 (which is the monoclonal antibody derived from the hybridoma identified with FERM BP4945) as labeled one. Details of sandwich method, one pot method, single antibody solid phase method, double antibody liquid phase method are described below. Sandwich method is a method described in the first specific aspect of the tenth aspect of the present invention, i.e., an immunoassay comprising at least the following steps of: (a) reacting sample with the immobilized monoclonal antibody or the immobilized antibody fragment of the present invention; and (b) reacting the labeled monoclonal antibody or the labeled antibody fragment with the antigen-antibody complex formed by binding human CETP in a sample with said immobilized monoclonal antibody or said immobilized antibody fragment. Based upon the present invention, most conventional methods by using enzyme or biotin as labeling substance specifically described below. Those methods are composed of such as the following steps but not limited to the following examples. Each of "immobilized monoclonal antibody" and "monoclonal antibody immobilized microplate" used below mean identical material. (Step 1) a step for preparing an immobilized monoclonal antibody(monoclonal antibody immobiOOOOlized microplate) by immobilizing the monoclonal antibody #72-1, #86-2 or #176-1 of the present invention which is reactive to human CETP on a microplate; (Step 2) a step to react a sample with the immobilized monoclonal antibody by adding a sample such as human blood plasma to the monoclonal antibody immobilized microplate; (Step 3) a step to remove non-reacted sample by washing the monoclonal antibody immobilized microplate; (Step 4) a step to prepare a labeled monoclonal antibody by labeling monoclonal antibody #72-1, #86-2 or #176-1 of the present invention which is reactive to human CETP with an enzyme such as biotin or peroxidase; (Step 5) a step to react labeled monoclonal antibody with an antigen-antibody complex (formed by reacting the immobilized monoclonal antibody with human CETP in a sample) by applying labeled monoclonal antibody to the monoclonal antibody immobilized microplate washed by "Step 3"; (Step 6) a step to remove non-reacted labeled monoclonal antibody from the antigen-antibody complex and by washing the monoclonal antibody immobilized microplate; (Step 7) a step to add avidin or enzyme modified avidin(in case biotin labeled monoclonal antibody is used at "Step 4"), or various substrates(in case enzyme labeled monoclonal antibody such as peroxidase labeled is used at "Step 4", depending on method used for measuring enzyme activity) together with a coloring agent(if necessary) to the monoclonal antibody immobilized microplate (washed at "Step 6) and to react with labeling substance on the labeled monoclonal antibody; (Step 8) a step to react avidin bound enzyme with substrate by adding various substrates depending on method for measuring enzyme activity in case enzyme modified avidin is used in "Step 7"; (Step 9) a step to stop the coloring reaction and enzymatic reaction by adding stop solution to the monoclonal antibody immobilized microplate; and (Step 10) a step to measure intensity of coloring, fluorescent or luminescence. Although the monoclonal antibodies #72-1, #86-2 and #176- 1 can be used both as immobilized form and as labeled form, it is preferable to use #72-1 as immobilized form and #86-2 and #176-1 as labeled form, more preferably #86-2. One pot method is an immunoassay mentioned in the first, the second or the third specific aspect of the tenth point of the present invention. The first is an immunoassay including at least the following steps of: (a) reacting sample with the immobilized monoclonal antibody or the immobilized antibody fragment of the present invention; and (b) reacting the labeled monoclonal antibody or the labeled antibody fragment with antigen-antibody complex formed by binding human CETP in a sample with said immobilized monoclonal antibody or said immobilized antibody fragment. The second is an immunoassay including at least the following steps of: (a) reacting a sample with the labeled monoclonal antibody or the labeled antibody fragment of the resent invention; and (b) reacting the immobilized monoclonal antibody or the immobilized antibody fragment of the present invention with the antigen-antibody complex formed by binding human CETP in a sample with said labeled monoclonal antibody or said labeled antibody fragment. The third specific aspect is the immunoassay comprising at least the following step of: (a) reacting a mixture comprising the immobilized monoclonal antibody or the immobilized antibody fragment of the present invention, the labeled monoclonal antibody or the labeled antibody fragment of the present invention and a sample. The above-mentioned three aspect are specifically explained hereinbelow by using examples. In the example, a step employs a generally used enzyme as a labeling agent, and comprises the following steps. However, the present invention is not limited to such examples given below. The terms "immobilized monoclonal antibody" and "monoclonal antibody immobilized beads" mean the same material. The first step is comprising the following steps. (Step 1) A step for producing immobilized monoclonal antibody(monoclonal antibody immobilized beads) by immobilizing monoclonal antibodies of the present invention having reactivity to CETP derived from human, #72-1, #86-2, and #176-1, on the beads respectively; (Step 2) a step for reacting an analyte in sample solution with a monoclonal antibody by adding both the monoclonal antibody immobilized beads and the sample solution such as human plasma together with buffer solution into a container having an internal space such as a test tube, plate or tube; (Step 3) a step for removing the solution in the container and washing the monoclonal antibody immobilized beads; (Step 4) a step for producing labeled monoclonal antibody by labeling monoclonal antibodies of the present invention, #72-1, #86-2, or #176-1, which capable of reacting with human CETP, with enzymes such as biotin or peroxides (Step 5) a step for reacting the labeled monoclonal antibody with antigen-antibody complex formed from reaction between immobilized monoclonal antibody and CETP derived from human by adding the labeled monoclonal antibody in the container containing the monoclonal antibody immobilized beads washed in Step 3; (Step 6) a step for removing an unreacted labeled monoclonal antibody which is not reacted with the complex with the solution in the container by washing the monoclonal antibody immobilized beads; (Step 7) a step for reacting a substance attached to the labeled monoclonal antibody with a substrate by adding either avidin or enzyme modified avidin when the biotin- labeled monoclonal antibody is employed in Step 4, or by adding a substrate suitable for enzyme activity assay systems employed when the monoclonal antibody labeled by enzymes such as peroxides is employed in Step 4, if desirably together with a color developing substance, in the container containing the monoclonal antibody immobilized beads washed in Step 6; (Step 8) a step for reacting an enzyme bound to avidin with a substrate suitable for the enzyme activity assay systems employed when the enzyme-modified avidin is added at Step 7; (Step 9) a step for terminating both enzyme reaction and color development by adding the terminator solution into either the reaction system of Step 7 or 8; and (Step 10) a step for determining the absorption intensity, fluorescence intensity or luminescence intensity. The second step is comprising the following steps. (Step 1) A step for producing labeled monoclonal antibody by labeling the monoclonal antibodies of the present invention having reactivity to CETP derived from human, #72-1, #86-2, and #176-1 with the labeling substance such as biotin and peroxidase. (Step 2) a step for reacting sample with labeled monoclonal antibody by adding both the labeled monoclonal antibody and the sample solution such as human plasma together with buffer solution into a container having the internal space such as the test tube, plate or tube; (Step 3) a step for producing an immobilized monoclonal antibody(monoclonal antibody immobilized beads) by immobilizing the monoclonal antibodies of the present invention having reactivity to CETP derived from human, #72-1, #86-2, or #176-1, onto the beads (-Step 4) a step for reacting the immobilized monoclonal antibody and the antibody-antigen complex formed from the labeled monoclonal antibody and the analyte, CETP derived from human in the sample by adding the monoclonal antibody immobilized beads in the reaction system of Step 2; (Step 5) a step for removing the solution in the container and unreacted labeled monoclonal antibody which is not reacted with the complex by washing the monoclonal antibody immobilized beads; (Step 6) a step for reacting the labeling substance attached to the labeled monoclonal antibody with the substrate by adding either avidin or enzyme modified avidin when the biotin-labeled monoclonal antibody is employed in Step 1, or by adding the substrate suitable for the enzyme activity assay systems when the monoclonal antibody labeled by enzymes such as peroxides*is employed in Step 1, in desirably together with the color developing substance, in the container containing the monoclonal antibody immobilized beads washed in Step 5; (Step 7) a step for reacting an enzyme bound to avidin with a substrate suitable for the enzyme activity assay systems employed added when the enzyme-modified avidin is added at Step 6; (Step 8) a step for terminating both enzyme reaction and color development by adding the terminator solution into either the reaction system of Step 6 or 7; and (Step 9) a step for determining the absorption intensity, fluorescence intensity or luminescence intensity. The third step is comprising the following steps. (Step 1) A step for producing immobilized monoclonal antibody(monoclonal antibody immobilized beads) by immobilizing the monoclonal antibodies of the present invention having reactivity to CETP derived from human, #72-1, #86-2, and #176-1, onto the beads; (Step 2) a step for producing labeled monoclonal antibody by labeling the monoclonal antibody having reactivity to CETP derived from human, #72-1, #86-2, and #176-1, with the enzyme such as biotin or peroxides; (Step 3) a step for simultaneous reacting the sample with both immobilized monoclonal antibody and labeled monoclonal antibody by adding the sample, both the monoclonal antibody immobilized beads produced in Step 1, the labeled monoclonal antibody produced in Step 2, and the sample solution such as human plasma together with the buffer solution into the container having the internal space such as the test tube, plate or tube; (Step 4) a step for removing the unreacted labeled monoclonal antibody which is not reacted with the complex by washing the monoclonal antibody immobilized beads the; (Step 5) a step for reacting the labeling substance attached to the labeled monoclonal antibody with the substrate by adding either avidin or enzyme modified avidin when the biotin-labeled monoclonal antibody is employed in Step 2, or by adding the substrate suitable for enzyme activity assay systems when the monoclonal antibody labeled with enzymes such as peroxides is employed in Step 2, in the container containing the monoclonal antibody immobilized beads washed in Step 4; (Step 6) a step for reacting an enzyme bound to avidin with a substrate suitable for the enzyme activity assay systems employed added when the enzyme-modified avidin is added at Step 5; (Step 7) a step for terminating both enzyme reaction and color development by adding the terminator solution into either the reaction system of Step 6 or 7; and (Step 8) a step for determining the absorption intensity, fluorescence intensity or luminescence intensity. In the above-mentioned concrete examples, all the monoclonal antibodies, #72-1, #86-2, and #176-1 may be employed as both the immobilized and labeled monoclonal antibodies. However, the monoclonal antibody #72-1 is preferable as the immobilized monoclonal antibody, and both the monoclonal antibodies #86-2 and #176-1 are preferable as the labeled monoclonal antibodies. The monoclonal antibody #86-2 is more preferable as the labeled one. A single antibody solid phase method is described above at the forth of the tenth concrete feature of the present invention, and it is an immunoassay comprising at least the following step of: (a) reacting a sample and a labeled CETP standard labeled with a labeling substance capable of providing a detectable signal independently or by reaction with other substances with the immobilized monoclonal antibody or the immobilized antibody fragment of the present invention. According to the present invention, the procedure which employs the labeling substance, particularly commonly used enzyme or biotin as the labeling substance, is explained in detail hereinbelow. For instance, the procedure comprises the following steps, however, the present invention is not limited to the example. Furthermore, the terms of "the immobilized monoclonal antibody" and "the monoclonal antibody immobilized microplate" has the same meaning. (Step 1) A step for producing immobilized monoclonal antibody(monoclonal antibody immobilized beads) by immobilizing the monoclonal antibodies of the present invention having reactivity to CETP derived from human, #72-1, #86-2, and #176-1, onto the microplate; (Step 2) a step for producing labeled CETP standard derived from human by labeling the CETP standard with the enzyme such as biotin or peroxides; (Step 3) a step for competitively reacting both the sample such as human plasma and the CETP standard with immobilized monoclonal antibody in the monoclonal antibody immobilized microplate; (Step 4) a step for removing the unreacted labeled CETP standard by washing the monoclonal antibody immobilized microplate; (Step 5) a step for reacting the labeling substance bound to CETP standard with a substrate by adding avidin or enzyme modified avidin as the substrate when the biotin labeled CETP standard derived from human is employed in Step 2, or a substrate suitable for enzyme activity assay systems employed when the enzyme-labeled CETP standard labeled by enzymes such as peroxides is employed in Step 2, if desirably together with a color developing substance in the monoclonal antibody immobilized microplate washed in Step 4; (Step 6) a step for reacting the enzyme bound to avidin with the substrate suitable for the enzyme activity assay systems when the enzyme-modified avidin is added at Step 5; (Step 7) a step for terminating both enzyme reaction and color development by adding the terminator solution into the monoclonal antibody immobilized microplate; and (Step 8) a step for determining the absorption intensity, fluorescence intensity or luminescence intensity. The monoclonal antibody #72-1, #86-2, and #176-1 may be used as the immobilized monoclonal antibody, however, the monoclonal antibody #72-1 is preferable. A double antibody solid phase method is described above at the fifth of the tenth concrete feature of the present invention, and it is an immunoassay comprising at least the following step of (a) or the following steps of (b) and (c): (a) reacting the monoclonal antibody or antibody fragment with a mixture of a sample and the labeled human CETP standard. The labeled human CETP standard is prepared by labeled labeling with a substance capable of providing detectable signal independently or by reaction with other substances; (b) reacting the monoclonal antibody or the antibody fragment of the present invention with a sample; (c) followed by the step (b), reacting the labeled human CETP standard with the reaction mixture of step (b). More specifically, the present invention is the immunoassay comprising at least the following steps of (a) and (d), or steps of (b) to (d). (a) reacting the monoclonal antibody or the antibody fragment of the present invention with a mixture of a sample and the labeled human CETP standard. The labeled human CETP standard is prepared by labeling with a substance capable of providing detectable signal independently or by reaction with other substance; (b) reacting the monoclonal antibody or the antibody fragment of the present invention with a sample; (c) followed by the step (b), reacting the labeled human CETP standard with the reaction mixture of step (b). The labeled human CETP standard is prepared by labeling with a substance capable of providing detectable signal independently or by reaction with other substance; (d) reacting an anti-serum derived from mammals reactive to said monoclonal antibody or said antibody fragment with an antigen-antibody complex. The antigen- antibody complex is formed by binding human CETP in said sample or said labeled human CETP standard with said monoclonal antibody or said antibody fragment. According to the present invention, the procedure which employs the labeling substance, particularly commonly used enzyme or biotin as the labeling substance is explained in detail hereinbelow. For instance, the procedure comprises the following steps, however, the present invention is not limited to the example. (Step 1) A step for producing labeled CETP standard derived from human by labeling the CETP standard with the enzyme such as biotin or peroxides; (Step 2) a step for competitively reacting (1) the mixture comprising both the sample such as human plasma and the CETP standard derived from human prepared in the above Step 1 to add into the container having internal space such as the test tube, plate, or tube to react with subsequently added the monoclonal antibody having the reactivity to the CETP derived from human of the present invention, #72-1, #86-2, or #176-1; (2) the sample such as human plasma is added into the container having internal space such as the test tube, plate, or tube to react with subsequently added the monoclonal antibody, #72-1, #86-2, or #176-1, having the reactivity to the CETP derived from human of the present invention; (Step 3) a step reacting the monoclonal antibodies, #72-1, #86-2, and #176-1, the sample added in Step 2, and the labeled human CETP standard added later when the labeled human CETP standard is not added simultaneously; (Step 4) a step for precipitating as immune-complex agglutinately by adding animal anti-sera except mice having the reactivity for mouse monoclonal antibody such as anti- mouse y globulin goat anti-sera to react the antigen- antibody complex of the monoclonal antibodies prepared in Step 2 to 3, #72-1, #86-2, or #176-1, and either human CETP in the sample or the labeled human CETP standard to form the complex comprising the antigen-antibody complex and the animal anti-sera except mice; (Step 5) a step for removing the agglutinate complex precipitated by centrifugation of the reaction system in Step 4; (Step 6) a step for reacting the immune-complex separated in Step 5 with labeling substance which is bound to avidin with either avidin or enzyme-modified avidin when the CETP standards are employed in Step 1, or the substrate suitable for the enzyme activity assay systems when the human CETP standard labeled by enzymes such as peroxides is used in Step 1, if desirably together with the color developing substance; (Step 7) a step for reacting the enzyme bound to avidin with the substrate suitable for the enzyme activity assay system when enzyme-modified avidin is employed in Step 6; (Step 8) a step for terminating both enzyme reaction and color development by adding the terminator solution into the monoclonal antibody immobilized microplate; and (Step 9) a step for determining the absorption intensity, fluorescence intensity or luminescence intensity. The monoclonal antibody #72-1, #86-2, and #176-1 may be used in the above-mentioned examples, however, the monoclonal antibody #72-1 is preferable. The term "affinity chromatography" in the present invention means the chromatography for separation or purification of human CETP contained in a sample by using the affinity between the antigen and antibody. As the examples of the sample, body fluids such as plasma, culture supernatants, or cetrifugation suprenatants are given. Specifically, the following two methods are given as examples. (1) A chromatography for separating the human CETP in the sample, wherein the sample is applied to the above- mentioned insoluble carrier such as filter or membrane for immobilizing the monoclonal antibody having reactivity to human CETP or its fragment of the present invention to separate the human CETP. (2) A chromatography for separating or purifying the human CETP in the sample, wherein the sample is applied to the above-mentioned insoluble carrier for immobilizing the monoclonal antibody or its fragment, and the carrier is packed into the column for applying the sample to elute to separate or purify the human CETP. In this method, as the example of the insoluble carriers, cellulose type, agarose type, polyacrylamide type, dextran type, polystyrene type, polyvinyl alcohol type, polyamino acid type ones and so forth are given. The monoclonal antibody or its fragment are immobilized on the insoluble carrier by means of the known method such as physical adsorption, polymerization by cross-link, trapping in the carrier matrix, or immobilization by non-covalent bond. Such carriers are packed into the columns such as a glass, plastic and stainless column having the configuration such as cylindrical one. Particularly, the latter chromatography is referred to as the affinity chromatography here. As the insoluble carriers for the affinity chromatography, any types of those may be used when they can immobilize the monoclonal antibody or its fragment of the present invention on them. As the examples, commercially available carriers such as Sepharose 2B, Sepharose 4B, Sepharose 6B, CNBr-Sepharose 4B, AH-Sepharose 4B, CH-Sepharose 4B, Activated CH-Sepharose 4B, Epoxy- activated Sepharose 6B, Activated thiol-Sepharose 4B, Sephadex, CM-Sephadex, ECH-Sepharose 4B, EAH-Sepharose 4B, NHS-activated Sepharose, Thiopropyl Sepharose 6B, and so forth(Pharmacia); Bio-gel A, Cellex, Cellex AE, Cellex-CM, Cellex PAB, Bio-gel P, Hydrazide Bio-Gel P, Aminoethyl Bio- Gel P, Bio-Gel CM, Affi-Gel 10, Affi-Gel 15, Affi-Prep 10, Affi-Gel Hz, Affi-Prep Hz, Affi-Gel 102, CM Bio-Gel A, Affi-Gel heparin, Affi-Gel 501, or Affi-Gel 601, and so forth(Bio-Rad); Chroma-Gel A, Chroma-Gel P, Enzafix p-Hz, Enzafix P-SH, Enzafix P-AB, and so forth(Wako Pure Chemicals); Ae-Cellulose, CM-Cellulose, PAB Cellulose, and so forth(Serva) are given. "Mice introduced DNA encoding human CETP" may be produced by the techniques employed generally in the field of the transgenic mouse production (See The newest animal cell experiment manual, AIC press, Chapter 7, pp. 361-408, 1990). Tall et al. reported that they produce a transgenic mouse introduced human CETP gene. However, the mouse can not secrete CETP in blood when it requires extrinsic or artificial induction such as taking foods supplemented zinc. In contrast, the transgenic mouse of the present invention secretes human CETP constantly without any induction described above, and such a transgenic mouse is not reported. "Pharmaceutical composition" contains the monoclonal antibody or its fragment of the present invention, and may contain pharmaceutically acceptable carriers such as excipients, diluent, vehicles, disintegrators, stabilizers, preservatives, buffering agents, emulsifiers, aromatics, coloring agents, sweetening agents, thickning agents, fravering agents, solubilizing agents, or other additives. Such pharmaceutical preparations may be formed as tablets, pills, powders, granules, injections, liquid preparations, capsules, troches, elixirs, suspensions, emulsions, or syrups. The pharmaceutical preparations may be administrated via oral or parental. In particular, the injections may be prepared by dissolving or suspending the pharmaceutical preparations in the pharmaceutically acceptable carrier without toxicity at the certain concentration. The carrier may be physiological saline, distilled water for injections. The certain range is from 0.1 µg of the monoclonal antibody/ml of carrier to 10 mg of the antibody/ml of carrier. Such injections may be administrated to patients need treatment at the dosage of 1 µg to 100 mg/kg of body weight, preferably at 50 µg to 50 mg/kg of body weight for one to several times per day. This administration is performed via clinically suitable routes such as intravenously, subcuteneously, intradermally, in intramuscle, in intraperitoneal and so forth. Preferable is intravenous administration. The present invention may be applied to not only hyperlipidemia but also atherosclerosis caused by the abnormal kinetics of CETP, prevention and/or treatment of various diseases such as hyperalphalipoproteinemia or hypercholes terinemia. (Examples) The present invention was explained in detail hereinbelow based on the following working Examples. However, the present invention is not limited to such examples. (Example 1) Establish of CETP activity assay system In the present invention for determining the CETP activity (cholesterol ester (abbreviated as CE) transporting activity), the assay system constructed as described below was employed unless it was specially stated. The assay system of the present invention is a modified one according to the method of Alberts et al., Arteriosclerosis, vol. 4, 49-58(1984). Briefly, the assay system was as follows. Firstly, a donor lipoprotein comprising high density lipoprotein in which radio-labeled cholesterol ester (HDL3) was incorporated. An acceptor lipoprotein comprising low density lipoprotein (LDL), and a sample containing CETP for assay were mixed to physiologically react with each other. Then, the amount of labeled CE transported from the donor lipoprotein to the acceptor lipoprotein was determined by using either the decrease of the radioactivity of the donor lipoprotein or increase of that of the acceptor lipoprotein to determine CETP activity of CETP in the sample. Preparation of donor lipoprotein Potassium bromide(KBr) was added to 20 ml of healthy volunteer plasma to adjust its comparative gravity, d=1.125 g/ral. After that, the plasma was subjected to density gradient centrifugation by 227,000 x g at 4°C for 17 hours, and then obtained the fraction of which comparative gravity was d>1.125 g/ml(HDL3 fraction). The fraction obtained was dialyzed against TBS(0.15 M NaCl/10 mM Tris(pH 7-4)). Then, 10 nM of tritium labeled cholesterol([3H]C, specific activity 50.3 Ci/mM) was gradually dissolved in 95 % ethanol with stirring slowly. The solution was incubated for 18 hr at 37°C. During the incubation, tritium cholesterol ([3H]C) attached onto the surface of HDL3 was esterified by Lecithin Cholesterol acyltransferase(LCAT) to be converted to tritium labeled cholesterol ester([3H]CE). Then [3H]CE was incorporated into HDL3. KBr was added into the human plasma to adjust its comparative gravity, d=1.21 g/ml. After that, the plasma was subjected to density gradient centrifugation by 227,000 x g at 4°C for 20 hours, and then obtained the fraction of which comparative gravity was d fraction obtained was dialyzed against TBS described before, and then obtained HDL3([3H]CE~HDL3; comparative gravity is 1.125 Potassium bromide (KBr) was added to 100 ml of healthy volunteer plasma to adjust its comparative gravity, d=1.019 g/ml. After that, the plasma was subjected to density gradient centrifugation by 227,000 x g at 4°C for 20 hours, and then obtained the fraction of which comparative gravity was d>1.019 g/ml. The fraction obtained was dialyzed against TBS described above. Then, KBr was added again to adjust the gravity d=1.063 g/ml. The solution was subjected to the density gradient centrifugation by 227,000 x g at 4°C for 20 hr, and then obtained the fraction with d TBS described above, and then obtained the fraction comprising LDL(the comparative gravity:1.019 The donor lipoprotein obtained in Example 1>( [3H]CE-HDL3 which contains 0.21 jig of cholesterol), the acceptor lipoprotein(LDL which contains 21 µg of cholesterol), and a sample for assay were mixed in a microtube. Then, TBS described above was also added into the microtube to adjust the ratio of cholesterol content in the donor lipoprotein to that in the acceptor lipoprotein equals to 1:100, and the ratio of cholesterol in HDL contained in the assay sample to that in the donor lipoprotein equals at least 1:10(total volume is 600 µl/tube). The tube was incubated for 15 hr in water bath at 37°C. Then, the tube was transferred onto ice for 15 minutes. After that, 400 µl of the iced TBS and 40 µ1 of 1 % dextran sulfate solution containing 0.5 M MgCl2 were added into the microtube, and vigorously vortexed. The microtube was incubated for 30 minutes on ice. Then, the tube was subjected to centrifugation by 8,000 x g at 4°C for 10 minutes, and then recovered HDL rich supernatant. The radioactivity in the supernatant was measured by means of a Scintillation counter. In order to determine CETP activity of the assay sample contained CETP, the radioactivity of the control sample which does not contain the assay sample was also measured by treating as the same as the procedure described above. CETP activity was determined based on the decrease of the radioactivity by comparing to that in the assay samples to that in the control sample. The activity was expressed as unit, and the CETP activity for transporting 1 nM of cholesterol per one unit time was represented as 1 unit(U). (Example 2) Preparation of purified CETP derived from human The procedure described below was performed at 4°C or in ice bath according to the known method. Dextran sulfate treatment The peripheral blood was obtained from healthy volunteers. Red blood cells in the blood were removed by the centrifugation by the known method, and then obtained 1 L of human plasma. Two liters of distilled water, 100 ml of 10 % dextran sulfate having molecular weight of 500,000 Da, and 80 ml of 4 M CaCl2 were added into the obtained plasma with gentle stirring on ice bath for 15 minutes. The mixed solution was centrifuged by 15,200 x g for 1 hr to fractionate insoluble dextran sulfate/lipoprotein complex as precipitate. The supernatant was collected, and 27.2 % BaCl2 was added to adjust the final concentration of the solution to 1.36 %. The mixed solution was stirred for 20 minutes, and then centrifuged by 15,200 x g for 1 hr. The precipitate was removed, and the supernatant was collected. Purification by phenyl Sepharose column chromatography Three molar of NaCl, 0.01 % sodium azide(NaN3), 50 pg/ml of Gentamysin sulfate, and 0.05 % ethylenediamine tetraacetic acid(EDTA) were added into the supernatant obtained in Example. The mixture was adjusted to pH 7.4. Phenyl Sepharose HP column(10 x 12.5 cm, Pharmacia Biotech) was equilibrated with 3 M NaCl(pH 7.4) containing 0.01 % NaN3, 50 µg/ml of Gentamysin sulfate, and 0.05 % EDTA as mentioned above. The mixture was applied into the phenyl sepharose column. After that the column was washed with 3 L of 0.15 M NaCl solution(pH 7.4), subsequently with 3 L of the solution without NaCl(pH 7.4), and then eluted with 20 % ethanol. The elution profile was monitored by using absorbance at 280 nm. The result was shown in Fig. 1. Based on the elution profile, active fractions containing CETP derived from human was collected. Purification by means of Resource Q column choromatography The active fractions collected in Example was dialyzed against the buffer(pH 7.4) comprising 25 mM NaCl and 10 mM Tris-HCl. Then, the dialyzed fraction was applied to Resource Q column(3.5 x 10 cm, Pharmacia Biotech) equilibrated with buffer. The column was washed with 300 ml of the above-mentioned buffer, and eluted with 2 L of 25-250 mM gradient NaCl(pH 7.4). The elution profile was monitored by absorbance at 280 nm. The result is shown in Fig. 2. Based on the elution profile, active fractions containing CETP derived from human was collected to obtain purified CETP derived from human. (Example 3) Preparation of anti-human CETP monoclonal antibody Human CETP monoclonal antibody was prepared according to the known method described in references such as Experimental Medicine suppl. "Cell Technology Handbook" (Toshio Kuroki et al. ed., pp. 66-74, Yodo press(1992)) or An introduction of monoclonal antibody experimental procedure(Tamio Ando et al., Kodansha(1991)). Immunization, cell fusion, and cloning Purified human CETP prepared in Example 2 with Freund"s complete adjuvant was injected into foot pads of BALB/c mice (female, 4-5 week age, purchased from Shizuoka experimental animal center) for the first immunization. Day 5th, 10th, 15th from the first immunization, the purified human CETP was injected into the foot pads of the immunized mice as the additional immunization. Furthermore, the purified human CETP was given similarly to the mice as the final immunization before one or two days from the day that the preparation of monoclonal antibody producing hybridoma was performed. Lymph nodes of knee was excised from the mice by a surgical operation according to known methods. The lymphocytes obtained from the lymph node and mouse myeloma cells, PAI were fused by using polyethylene glycol 4000 as a fusion agent (at the ratio of 3:1). Then, the hybridomas were cultured in ASF104 medium(AGF) comprising HAT with aminopterine containing 10 % fetal calf serum. Then the media was changed to HT medium without aminopterine to select the hybridoma prepared by fusion of the lymphocytes and the myeloma cells. The number of obtained clones was 169. Screening of hybridoma Hybridoma producing anti-human CETP monoclonal antibody was screened by the CETP activity assay system established in Example 1. The purified human CETP prepared in Example 2 and each culture supernatant of the hybridoma screened(60 µl) were added into the mixture of the donor lipoprotein prepared in Example and the acceptor lipoprotein prepared in Example containing 21 µg of cholesterol). Then, TBS was added to prepare the total volume of 600 µl/tube. The prepared solution was treated similarly to the procedure described in Example was determined in each sample by comparing with the activity of the control sample to obtain the CETP inhibition activity(inhibition activity against CE- transporting activity of CETP) in the culture supernatant of the hybridoma. By means of the screening, three positive clones, #72-1, #86-2, and #176-1, which have different CETP inhibition activities were obtained. Two hybridoma, #72-1 and #86-2 were deposited to National Institute of Bioscience and Human-Technology Agency of Industrial Science and Technology at the date of Dec. 20, 1994, and International accession numbers are FERM BP-4944 and FERM BP-4945, respectively. Large scale preparation of monoclonal antibody Each of the hybridoma #72-1, #86-2, or #176-1 (106 to 107 cells/0.5 ml/mouse) were administrated to 15 of ICR nude mice(female, 7-8 week age, purchased from Charles River) via intraperitoneally. Ten days later, ascites were collected from the mice anesthetized according to the known method. The monoclonal antibodies were prepared in large scale as supernatants of centrifugation. Isotyping Isotyping for the monoclonal antibodies, #72-1, #86-2, and #176-1, were performed with Mouse monoclonal antibody isotyping kit(Amersham) according to the protocol supplemented to the kit. The result showed that both #72-1 and #86-2 were IgGl, and #176-1 was IgG2b. Purification of the monoclonal antibody Five ml of the each monoclonal antibodies, #72-1, #86- 2, and #176-1, prepared as the supernatant, were diluted by triple times with 0.06 M acetate buffer(pH 4.0), and pH was adjusted to 4.8 with 1 N HC1. Then, 16.5 pi of caprilyc acid(Wako Pure Chemical) was gradually added into the diluted solution at room temperature to react for 30 minutes. After that, the mixture was centrifuged by 10,000 rpm for 20 minutes to precipitate proteins except antibodies. Supernatants were collected, and filtrated with the filter(Millipore) to remove white precipitates. Filtrates were dialyzed against phosphate buffer for 2 hr. After dialysis, ammonium sulfate(26.2 g/100 ml) was added little by little with stirring to react at 4°C for 120 minutes. Then, the mixture was centrifuged by 10,000 rpm for 20 minutes to collect precipitates. Phosphate buffer was added to the precipitate, and dialyzed against to phosphate buffer at 4°C for 24 hr, and then obtained the purified monoclonal antibody, #72-1, #86-2, and #176-1, respectively. (Example 4) Purification of human CETP by affinity chromatography The purified human CETP prepared in Example 2 was further purified by means of the affinity column chromatography which used the monoclonal antibody, #72-1, and then characterized. Preparation of immobilized monoclonal antibody as a adsorbent media for column packing HiTrap-NHS-activated Sepharose HP column(1 ml, Pharmacia Biotech) was employed, and the experiment operation was performed according to the attached protocol. The monoclonal antibody, #72-1, was dissolved in 0.2 M sodium hydrogen carbonate(pH 8.3) containing 0.5 M NaCl was injected into the column at the ratio of 5 mg/ml. Then, the monoclonal antibody was reacted with the column for 45 minutes to immobilize onto NHS-activated Sepharose to prepare the immobilized monoclonal antibody #72-1. Purification by affinity chromatography A solution containing the purified human CETP obtained in Example 2 was applied on the column prepared in Example. Then, the column was washed with 5 ml of 50 mM Tris-HCl buffer(pH 7.6) containing 500 mM NaCl, and eluted with 2 mM glycine-HCl(pH 2.8). The eluted fraction was neutralized with 2 M Tris-HCl(pH 8.8). The elution profile was monitored by absorbance at 280 nm. The result was shown in Fig. 3. Based on the elution profile, active fractions containing human CETP were pooled. The pooled fraction were dialyzed against 50 mM Tris-HCl containing 0.15 M NaCl(pH 7.6), and the highly purified human CETP was prepared. Characterization of purified human CETP As described in Fig. 3, enzyme activity (specific activity) of CETP obtained in Example 4 was 1,100 U/mg. The molecular weight of the purified human CETP was analyzed by using SDS-PAGE(sodium dodecyl sulfate polyacrylamido gel electrophoresis). The result was shown in Fig. 4. It was confirmed that the purified human CETP obtained in Example 4 comprises two proteins with about 63,000 Da and 61,000 Da. Many researchers tried to purify the human CETP, and the values of the molecular weight were reported. Such values were not identical each other, because the purification method, purity of the protein and/or analysis of the molecular weight were different. However, they are in the range of about 64,000 to 66,000 Da. Compared to such values, it was demonstrated that the purity of the CETP obtained in Example 4 was very high. (Example 5) Characterization of Anti-human CETP monoclonal antibody Reactivity to the human CETP The reactivity of each of the purified monoclonal antibodies, #72-1, #86-2, and #176-1, prepared in Example 3 was confirmed by determining human CETP transporting activity assayed by means of CETP activity assay system established in Example 1 as the same as Example. The purified human CETP prepared in Example 4, and each of the monoclonal antibodies, #72-1, #86-2, and #176- 1, which were diluted with Tris-HCl buffer(pH 7.4) containing 0.15 M NaCl at the various concentration, were added into the mixture of the donor lipoprotein([3H]CE-HDL3 containing 0.21 g of cholesterol) prepared in Example and the acceptor lipoprotein(LDL containing 21 ^g of cholesterol) prepared in Example further added to prepare the total volume of 600 µl/tube. The prepared solution was treated similarly to the procedure described in Example each sample and the control without the monoclonal antibody were determined. Comparing the values of the corresponding samples, the CETP inhibition activity of each monoclonal antibody was determined. As the reference, mouse IgG was also used. The result is shown in Fig. 5. The monolonal antibody, #72-1, showed dose-dependent inhibition of the human CETP activity, particularly, the human CETP activity was inhibited 100 % at the concentration equal to or more than 10 µg/ml. In contrast, the monoclonal antibody, #86-2, did not show dose-dependent inhibition. This monoclonal antibody showed the maximum inhibition(58 %) at the concentration about 0.5 /jg/ml; however, it did not show any inhibition at the concentration equal to or more than 17 µg/ml. The monoclonal antibody, #176-1, did not show dose-dependent CETP inhibition activity, neither did #86-2, and it showed the maximum inhibition(50 %) at the concentration of about 1.0 ^g/ml. Based on the above results, it was demonstrated that the epitope recognized by the monoclonal antibody #72-1 was different from that recognized by other monoclonal antibodies, #86-2 or #176-1 with high possibility. Reactivity for rabbit CETP The respective reactivity of the monoclonal antibodies, #72-1, #86-2, and #176-1 prepared in Example 3 for the rabbit CETP was confirmed by means of the CETP activity assay system similar to that of Example 1. The purified rabbit CETP, the donor lipoprotein and the acceptor lipoprotein used for the assay system were prepared as described below. Preparation of the purified rabbit CETP The following procedure was performed according to the known method at 4°C or in ice bath. (1) Dextran sulfate treatment Peripheral blood was obtained from plural rabbits(Japan white, Kitayama rabes). The blood was centrifuged to remove red blood cells according to the conventional method, and one hundred ml of rabbit plasma was obtained. Two hundred ml of distilled water, 10 ml of 10 % of dextran sul fate (molecular weight 500,000 Da), and 7.9 ml of 4 M CaCl2 were added into the 100 ml of the plasma obtained, and gently stirred in ice bath for 15 minutes. The mixture was centrifuged by 20,000 x g for 20 hr at 4 °C to fractionate the insoluble dextran sulfate/lipoprotein complex, and then collected the supernatants. (2) Purification by Phenyl Sepharose column chromatography Three molar of NaCl, 0.01 % NaN3, 50 µg/ml of Gentamysin sulfate, and 0.05 % EDTA were added to the supernatant obtained, and adjusted pH 7.4. Prior to apply the mixture, Hiload 26/10(Phenyl Sepharose column, Pharmacia Biotech) was equilibrated with the same solution described above. Then, the mixture was applied on the column, and washed with 0.15 M NaCl solution (pH 7.4) containing the same concentration of NaN3, Gentamysin sulfate and EDTA. After that, adsorbed components were eluted by using the solution without NaCl and 20% ethanol. The elution profile was monitored absorbance at 280 nm. The result is shown in Fig. 6. Based on the elution profile, the active fractions containing rabbit CETP were pooled. (3) Purification of Blue Sepharose column chromatography The active fractions pooled in the above (2) was dialyzed against 50 mM HEPES buffer(N-2- hydroxyethylpiperadine-N" -2-ethanesulfonic acid) (pH 7.0). Then, the dialyzed sample was applied on Blue Sepharose column(CL-6B, 1.5 x 8.5 cm, Pharmacia Biotech) which was equilibrated with the buffer. The column was washed with the buffer, and then eluted with the solution containing 1.5 M NaCl and the buffer(pH 7.0). The elution profile was monitored by absorbance at 280 nm. The result is shown in Fig. 7. Based on the elution profile, the active fractions containing rabbit CETP were pooled. (4) Purification by succinylated LDL Sepharose column chromatography The active fractions pooled in the above (3) was dialyzed against the 50 mM HEPES buffer(pH 7.4). Then, succinylated LDL Sepharose column was prepared by using succinyl LDL and CNBr-activated Sepharose 4B(1.5 x 8.5 cm, Pharmacia Biotech). The column was equilibrated with the 39 mM phosphate buffer containing 0.025 % EDTA and 60 mM NaCl(pH 7.4). Then, the dialyzed sample was applied on the column. The column was washed with the same buffer, and then eluted with the buffer(pH 7.4) containing 0.01 % EDTA. The elution profile was monitored by absorbance at 280 nm. The result is shown in Fig. 8. Based on the elution profile, the active fractions containing rabbit CETP were pooled. The donor lipoprotein and the acceptor lipoprotein According to the similar method described in both Example HDL3 containing 0.21 µg cholesterol) and the acceptor lipoprotein(LDL containing 21 µg of cholesterol) were prepared by using the rabbit plasma. Confirmation of the reactivity for rabbit CETP The respective reactivity of the monoclonal antibodies, #72-1, #86-2, and #176-1 against the rabbit CETP was confirmed by means of the CETP activity assay system in which purified rabbit CETP prepared in , and the donor lipoprotein prepared and the acceptor lipoprotein prepared in were used as the same as that of Example . The result is shown in Fig. 9. Neither monoclonal antibody #86-2 nor #176-1 showed rabbit CETP inhibition activity independent of their concentration. Accordingly, these monoclonal antibodies did not have the specific reactivity for the rabbit CETP. Furthermore, the monoclonal antibody #72-1 did not show the detectable reactivity at the concentration equal to or less than 3 fig/ml. (Example 6) Preparation of antibody fragments, F(ab")2 and Fab The F(ab")2 and Fab of the purified monoclonal antibodies prepared in Example 3, #72-1, #86-2, and #176-1 were prepared as follows. Each of the monoclonal antibodies was added into 20 mM acetate buffer(pH 3.5) at the concentration of 5 mg/ml respectively, and incubated for 30 minutes. Then, one ml of insolubilized pepsin (Pierce) was added to the incubated solution, and further incubated for 12 hr at 37°C rotating by using a rotator. After that, the reaction mixture was centrifuged by 3,000 rpm for 10 minutes at 4 °C to collect supernatants as the first supernatant. The supernatant was subjected to the Protein A affinity chromatography by using Protein A column kit (Amersham), and the procedure was performed acceding to the attached protocol. A binding buffer was added to the precipitate to suspend, and then the suspension was centrifuged by 3,000 rpm for 10 minutes at 4 °C to collect supernatants as the second supernatant. Both the first and second supernatant were pooled, and the same amount of the binding buffer was added. Then, 1 N sodium hydroxide was added to adjust pH 8.9. Protein A Sepharose column was equilibrated with the binding buffer prior to applying the mixture. The column was washed twice with 5 ml of the binding buffer, and then the eluted fractions were obtained. The eluted fractions were dialyzed against 2 L of 5 mM phosphate buffer pH 6.8 for 24 hr at 4°C. Further purification, the dialyzed sample was subjected to HPLC by using Hydroxyl apatite column(Bio Rad). The dialyzed sample was applied onto the Hydroxyl apatite column. Then, the column was washed with 5 mM phosphate buffer for 5 minutes. Retained components in the column were subsequently eluted, with gradient elution buffer of 5 mM to 0.4 M phosphate buffer. The eluents were fractionated by using a fraction collector. The elution profile were monitored by absorbance at 280 nm to monitor the amount of F(ab")2 contained in the fracitions to pool the active fractions. The pooled fractions were dialyzed against 2 L of phosphate buffer at 4 °C for 24 hr to obtain the respective F(ab")2 of the monoclonal antibodies, #72-1, #86-2, and #176-1. (Example 7) Establishment of human CETP assay method by using sandwich ELISA Preparation of the solid phase with immobilized monoclonal antibody Each of the purified monoclonal antibodies, #72-1, #86-2 and #176-1 prepared in Example 3 were diluted with phosphate buffer(pH 7.4) so as to be 10 ng/well to 1 µg/well. Fifty pi of the diluted antibody solutions were added into wells of 96 well microtiter plates for ELISA(Corning). The plates with the solutions were incubated at 4°C for 24 hr to adsorb each monoclonal antibody onto the inside of the well. Then, the well was washed with 300 jjI of phosphate solution containing 0.1 % Tween 20 for four times. After discarded the phosphate buffer, 300 µl of the blocking agent, Block Ace(Dainippon Pharmaceutical), was added to the well to incubate for 2 hr to block the antibody unbound sites. Each well was washed with 300 µl of phosphate solution containing 0.1 % Tween 20 for four times. Preparation of labeled monoclonal antibodies One mg/ml of each of the purified monoclonal antibody solutions, #72-1, #86-2 and #176-1, prepared in Example 3 were dialyzed against 0.1 M NaHCO3(pH 8.2 to 8.3) at 4 °C for 24 hr. Then, one hundred pi of NHS-biotin(2 mg/ml, Funakoshi) was added into the dialyzed antibody solution, and stirred vigorously. The antibody solution was incubated for 4 hr at room temperature. After that the antibody solution was dialyzed against phosphate buffer at 4 °C for 4 hr. Establishment of the assay method by using sandwich ELISA The human CETP assay method by using sandwich ELISA established of the present invention was as follows. Each of the monoclonal antibodies, #72-1, #86-2, and #176-1 was immobilized respectively onto the microplates at the concentration of 10 ng/well to 1 ^g/well as described in . The above-mentioned microplates prepared was referred to as the immobilized microplate. The immobilized plates were washed with phosphate buffer containing 0.1 % Tween 20 for 3 times. Phosphate buffer containing 10 % Block Ace was used for diluting the samples which were the purified human CETP standard prepared in Example , the plasma of healthy volunteers without lipoprotein prepared in Example , or the plasma of various patients. The immobilized plates with 100 µl of the diluted samples were incubated at room temperature for 1 hr. The plates were washed with the phosphate buffer containing 0.1 % Tween 20 for three times. The biotin labeled monoclonal antibodies, #72-1, #86-2, or #176-1, prepared in Example were diluted with the phosphate buffer containing 10 % Block Ace to 3 ng to 1 µg/50 µl/well. Then, the plates with 50 µl of the biotin labeled monoclonal antibodies were incubated at room temperature for 1 hr. After that, the plates were washed with the phosphate buffer containing 0.1 % Tween 20 for 3 times. Fifty µl of streptoavidin-ß- galactosidase(Gibco BKL) diluted 1,000 times with 20 mM HEPES buffer containing 0.5 M NaCl(pH 7.0) was added into each well of the immobilized plates. Then, the immobilized microplates were incubated at room temperature for 1 hr. The immobilized microplates were washed with the phosphate buffer containing 0.1 % Tween 20 for three times. After that, fifty µ1 of 0.015 % or 0.01 % of 4-methyl- umbelliferyl-ß-D-galactoside(Sigma) diluted with 10 mM phosphate buffer containing 1 mg/ml of BSA, 100 mM NaCl, 1 mM MgCl2(pH 7.0) was added into each well of the immobilized microplates. Then, the immobilized microplates were incubated at room temperature for 10 or 20 minutes. After that, one hundred jvl of Na2CO3 solution was added into the wells to terminate the reaction. Intensity of the wells were measured with Fluoroscan II microplate fluorometer(Flow Laboratories) at 460 nm of the emission wave length and the 355 nm of excitation one. The amount of CETP in the sample was determined from the calibration curve obtained in the following Example. The results of the assay when various combination of the monoclonal antibodies(#72-1, #86-2, and #176-1) immobilized or labeled are shown in Figs. 10 to 12. As a result, it was confirmed that the above-mentioned monoclonal antibody amount were quantitatively determined when the different monoclonal antibodies immobilized or labeled were combined, for example, #72-1 and #86-2, or #72-1 and #176-1. The optimal concentration of the immobilized or the labeled monoclonal antibody were determined under the following conditions. As the substrate, 4-methyl- umbelliferyl-ß-D-galactoside was used, and as the assay sample, the plasma from the healthy volunteers without lipoprotein prepared in Example 2 was used. (1) the immobilized monoclonal antibody: #72-1(10 ng to 1 µg/well) the labeled monoclonal antibody: #86-2 or #176-1(10 ng to 1 µg/well) substrate concentration: 0.01 % reaction time: 10 minutes (2) the immobilized monoclonal antibody: #72-1(1 µg/well) the labeled monoclonal antibody: #86-2(1 to 3 ng/well) substrate concentration: 0.015 % reaction time: 20 minutes The result are shown in Figs. 13 to 17. As shown in Figs. 13 to 17, the optimal concentration of the immobilized monoclonal antibody, #72-1 was 1 jig/well(20 µg/ml). That of the labeled monoclonal antibody, #86-2 was 100 ng/well(2 µg/ml) when the reaction time was 10 minutes, and 3 ng/well(60 ng/ml) when the reaction time was 20 minutes. Accordingly, the optimal concentration of the labeled monoclonal antibody may be changed since the concentration depends on the reaction time for substrate. In the following Examples, the sandwich ELISA was employed, and the conditions were as follows: the immobilized monoclonal antibody: #72-1(1 µg /well (20 µg/ml)) the labeled monoclonal antibody: #86-2(3 ng/well(60 µg/ml)) the reaction time for substrate: 20 minutes Preparation of a calibration curve and confirmation of accuracy A calibration curve was prepared by using the sandwich ELISA established in Example . As Standard, the purified human CETP prepared in was used. As the immobilized monoclonal antibody, #72-1(1 µg/well), and as the labeled one, #86-2(3 ng/well) were also used. The result is shown in Fig. 18. As shown in Fig. 18, the calibration curve was almost linear at the range of very low concentration between 0.4 to 1.25 ng/ml(correlation coefficient: r=0.995). The determination limit (the detection limit) of the sandwich ELISA of the present invention was 0.4 ng/ml. Accordingly, it was demonstrated that the sandwich ELISA of the present invention had excellent sensitivity of determination, that is, excellent sensitivity of detection. Calculation of coefficient of variation In order to study the accuracy of the assay method of the present invention, the coefficient of variation(c.v.) among 48 wells in a 96 well plate or different 6 plates was calculated. The c.v. obtained in the same plate was 7.06 %, and that obtained in different plates was 6.13 %. Accordingly, it was demonstrated that the assay system by using the sandwich ELISA of the present invention has high accuracy. Preliminary experiment for establishment of the human CETP assay system Study of the effect of the determination(assay) capability by human plasma components In order to study whether the plasma components which interfere the human CETP determination was contained in human plasma or not, the following tests were performed. The purified human CETP prepared in Example with either 0.735 µg/ml, or 1.469 µg/ml were added into the plasma from the healthy volunteers prepared in Example 1> which contained 0.56 µg/ml of CETP. Then, The amount of CETP was determined by using the method established in the above-mentioned . When 0.735 pg/ml or 1.469 pg/ml of CETP was added, the total CETP amount of each sample before incubation was calculated as 1.294 µg/ml or 2.029 µg/ml respectively. On the other hand, the total amounts of CETP recovered were 1.440 µg/ml and 2.189 µg/ml, and the recovery percentages were 111 % and 108 % respectively. As a result, it was demonstrated that the human plasma did not contain any component which interfere the determination(assay) of CETP by the method of the present invention. Accordingly, it was shown that the amount of CETP in human plasma was accurately determined by using the method of the present invention. Effect of lipoproteins in human plasma In order to study effect given by the substances except CETP such as lipoproteins for the human CETP determination, the human CETP with different purity as the assay sample was determined by using the assay system established in the .above-mentioned . For the determination, the following samples were used as the samples. (1) the plasma without hemocytes by centrifugation prepared in Example (2) the plasma without lipoprotein finally prepared in Example (3) the purified CETP with the Phenyl Sepharose column prepared in Example (4) the purified human CETP prepared in Example The result is shown in Fig. 19. Dilution curves obtained were respectively parallel with that of the purified human CETP(4). As a result, it was demonstrated that the lipoproteins in human plasma did not interfere the determination of CETP by the method of the present invention. Accordingly, it was shown that the determination method of the present invention may be apply to plasma irrespective of the sample"s purity. Correlation of CETP contents and CETP activities In order to investigated the precision of the determination method of the present invention, the correlation between the determined CETP amount in the human plasma sample and the CETP activity of the plasma sample per se was investigated. CETP contents were determined by using the plasma without lipoproteins obtained in Example as the assay sample. CETP activity was measured by using the same plasma sample with the CETP assay system of Example 1. The result is shown in Fig. 20. Since correlation coefficient(r) obtained was 0.953, the amount of CETP determined by the method of the present invention and the CETP activities of the same sample show good correlation. As a result, it was demonstrated that the assay system of the present invention has quite high precision. (Example 8) Determination of the human CETP in human plasma The amounts of CETP of from the healthy volunteers or various patients were determined by using the sandwich ELISA method established in Example . The CETP activity of the plasma was determined by using the donor lipoprotein([3H]CE~HDL3 which contains 6 ^g of CE) prepared similar to Example and the acceptor lipoprotein (LDL which contains 600 yq of CE) prepared similar to Example established in Example The human plasma used in the assay were plasma from the healthy volunteers (13), and patients of type Ha hyperlipidemia(5), type lib hyperlipidemia(5), CETP deficiency(2), LCAT deficiency(2), and hyperaplha- lipoproteinemia(l). Results are shown in Fig. 21. Any sample employed in the assay showed the good correlation between the amount of CETP determined and CETP activity. As expected, CETP was not detected in the plasma form the patients of CETP deficiency. Furthermore, it was demonstrated that the assay method of the present invention may be applied on not only the plasma from healthy volunteer but also the patients with various diseases. (Example 9) Comparison with the conventional determination method The following tests were performed to study the availability of the assay system of the present invention comparing with the conventional method which employs sandwich ELISA. In the conventional method, treatments by detergents such as TritonX-100 or heating is performed to expose epitopes capped by the interaction between protein-protein or protein-lipoprotein prior to the determination. However, such treatment causes denaturation of the proteins such as CETP in the sample, and the protein loses its higher-order structure to become inactivate biologically. Since CETP detected or determined by the conventional method was not an intact one, but denatured one, it was impossible to determine the accurate amount of CETP in the plasma. Accordingly, the followings were used as the sample in this determination. (1) the plasma from healthy volunteer without lipoprotein prepared as the same as Example (2) the plasma from healthy volunteer without lipoprotein incubated with 0.1 % Tween 20 (3) the plasma from healthy volunteer without lipoprotein heated at 95°C for 5 minutes (4) the plasma from healthy volunteer without lipoprotein incubated with 1 % Tween 100 The result was shown in Fig. 22. CETP was essentially detected in all samples, (1) to (4), however, it was not detected in both samples, (3) and (4). Particularly, no CETP was detected in (3). Furthermore, CETP activity was decreased more than 30 % in the plasma treated with the low concentration of Tween 20. Namely, intact CETP in the human plasma was not detected by using the conventional determination(assay) method which requires the detergent treatment or heating. However, it could be determined precisely by using the assay system of the present invention, since the monoclonal antibody has the specific reactivity for intact CETP in the human plasma. Furthermore, it was also demonstrated that the known monoclonal antibodies for the human CETP react specifically to the denatured human CETP, but not react to human intact CETP. (Example 10) Preparation of the pharmaceutical compositions and their CETP inhibition activity The purified monoclonal antibodies, #72-1, #86-2, and #176-1 were dissolved in 10 ml of the distilled water for injections to prepare injections at the concentration of 50 to 150 µg/wl. Anti-rabbit CETP monoclonal antibody was prepared similarly to Example 3 by using the purified rabbit CETP prepared in Example as immunogen. Briefly, BALB/c mice(female, 4-5 weeks, Shizuoka Experimental Animal Center) was immunized with the immunogen to obtain two hybridoma #2-64 and #9-1, which produced anti-rabbit monoclonal antibody. Each hybridoma was transplanted into nude ICR mice(female, Charles River) to prepare anti-rabbit monoclonal antibodies from the ascites. The monoclonal antibodies obtained, #2-64 and #9-1 were both IgGl. The monoclonal antibody, #2-64 was dissolved into the distilled water for injections at the concentration of 0.13 mg/ml. The monoclonal antibody, #9-1, was at the concentration of 5.53 mg/ml. Thus prepared injections containing either #2- 64 or #9-1 was administrated into rabbit at the dose of 1.8 mg/kg of body weight via i.v. The time of initial administration was time 0, and the monoclonal antibodies were administrated as the same dose in every 24 hr. Blood was sampled at 1, 2, 4 hr from the initial administration to obtain plasma. The CETP activities in the plasma thus obtained were determined by using the sandwich ELISA for the rabbit CETP determination method established with these monoclonal antibodies, #2-64 and #9-1, similarly to Example 1. It was demonstrated that the monoclonal antibodies, #2-64 and #9-1, significantly inhibit CETP in rabbit plasma. (Example 11) Production of human CETP high expression transgenie mice The plasmid containing the cDNA encoding the human CETP was obtained as follows. A cDNA fragment encoding the human CETP was amplified by PCR with 5"-stretch cDNA library of human liver(Clonetech) as a template. The PCR products were separated by agarose gel electrophoresis with the established known method. The cDNA fragment of interest was excised from the gel followed by ligation kit(Takara). The ligated plasmid was introduced into E. coli DH5 to obtain enough amount of plasmid to perform DNA sequencing. After sequencing, the human CETP cDNA was confirmed. cDNA encoding the human CETP obtained above was treated with DNA ligation kit(Takara), and inserted into a vector containing human ß-actin promoter(Mol. Cel. Immunol., 4:1961-1969(1984); ibid., 5:2720-2732(1985)) to obtain the plasmid, pCETP-1. By electroporation, pCETP-1 was introduced into COS cells, and then obtained the RNA of transformed COS cells according to the known method. RT- PCR was performed by using the RNA to confirm the specific transcription of human CETP in COS cells introduced pCETP- 1. Another transformant was produced similarly to the COS cells by using the expression vector pME 18S. The expression of human CETP in the trans formant was assayed by using the sandwich ELISA established in Example 7. In order to produce transgenic mice, the plasmid, pCETP-1 was enzymatically treated so as to linear. ICR white mice having vaginal plug(female, Japan Crea) were employed as foster mice, . This mouse was obtained by mating a female ICR mouse with a male ICR mouse with seminal duct ligation(Japan Crea). C57BL/6J mice(female, Japan Crea) were employed as mice for collection of embryo for human CETP gene transduction. This mouse was obtained by mating male C57BL/6J mouse with a female C57BL/6J mouse given both 5 unit of PEAMEX and the same unit of PUBEROGEN(both from Sankyo) for superovulation. After mating, oviducts were excised from the female C57BL/6J mouse to be treated with hyaluronidase to obtain embryo alone to store in media. Introduction of human CETP gene into the embryo was performed by using a manupilater under a microscope according to the known method. The embryo was retained by holding pipette at 37°C. The solution containing the linear gene of human CETP prepared before was introduced into male pronucleus of the embryo by using injection pipette. After introducing the gene, embryos holding normal condition were selected to be introduced into the oviducts of the tentative parent mouse, ICR white mouse. Tails of progeny mice from the parent mouse, which were hetero mice, were cut off to investigate whether the human CETP gene was introduced into the mouse genomic gene. By using PCR method, this introduction into the mouse genomic gene was confirmed, and the expression of the human CETP was also determined by using the sandwich ELISA established in Example 7. Mating two hetero mice described above, the transgenie mice which highly express human CETP was produced as homo mice. (Example 12) In vivo CETP inhibition of anti-CETP monoclonal antibody Two purified human anti-CETP monoclonal antibodies, #72-1 and #86-2 prepared in Example 3 was dissolved in the distilled water for injections at the ratio of concentration 29:1 to prepare injections, respectively. The prepared solution was administrated i.p. to 3 of the female transgenic mice produced in Example 11 at the dose of 100 mg/kg as single injection.. These mice were fed pellet type food for breeding freely for 4 months. Just before the antibody administration was time 0, and blood was sampled at the time, 1, 3, 6, and 24 hr from eyegrounds to separate plasma by centrifugation. The CETP activity in the plasma obtained(CETP inhibition activity by anti-human CETP antibody) was determined as the amount of CE transportation from HDL to LDL by using the CETP activity determination system established in Example 1. The result is shown in Fig. 24. It was confirmed that the monoclonal antibody of the present invention completely inhibit the human CETP activity in vivo. Furthermore, such inhibition effect for the human CETP activity in vivo was firstly clarified by this invention. (Example 13) Prevention of atherosclerosis by anti-CETP monoclonal antibody Two purified human anti-CETP monoclonal antibodies, #72-1 and #86-2 prepared in Example 3 was dissolved in the distilled water for injections at the ratio of concentration 29:1 to prepare injections, respectively. The prepared solution was administrated i.p. to 3 of the female transgenic mice produced in Example 11 at the dose of 75 mg/kg per injection for 4 days. These mice were fed pellet type food for breeding freely for 6 weeks. PBS(Phosphate buffered saline) was administrated to other mice as control. Just before the antibody administreition was time 0. Blood was sampled at the day -3, 2, 4, 3, and 11 from eyegrounds to separate plasma by centrifugation. The amounts of cholesterol in the plasma obtained was determined by using lipoprotein determination kit(Liquitec TC1/TC2, Boehringer Mannheim). For the fractionation of HDL, HDL cholesterol separation reagent(Boehringer Mannheim) was employed. The result is shown in Fig. 25. It was demonstrated that HDL cholesterol level in plasma was significantly enhanced when the anti-human CETP monoclonal antibody of the present invention was administrated in vivo. HDL was considered to be an important protein having anti-arteriosclerosis effect. In fact, it was showed that the development of atherosclerosis legion was inhibited or recessed by enhancement of HDL in plasma(J. Clin. Invest., 85:1234-1241(1990)). The observation that the administration of anti-human CETP antibody in vivo enhances the HDL cholesterol level in plasma was clarified at first by the present invention. Accordingly, the result of the above-mentioned test revealed that the monoclonal antibody of the present invention was available for prevention and/or treatment of atherosclerosis. INDUSTRIAL UTILITY Since the anti-human CETP monoclonal antibody of the present invention has higher binding specificity(CETP inhibition activity) to human CETP, particularly intact CETP in the human body fluid such as plasma, the above- mentioned intact CETP per se may be determined convenient and highly sensitive. Such determination(assay) method has not been established before. Particularly, by using the above-mentioned characteristics of the any of three anti-human CETP monoclonal antibodies of the present invention, the intact CETP can be determined more convenient and sensitive when any of the two monoclonal antibodies described above in combination were employed in the sandwich ELISA. In the determination method employing the monoclonal antibody of the present invention, the amount of CETP in bodily fluid such as plasma is determined conveniently with high sensitivity without any pretreatment such as the detergent or heat treatment. Since the anti-human CETP monoclonal antibody of the present invention has excellent CETP inhibition activity, it was available as Pharmaceuticals for prevention and/or treatment for various diseases such as atherosclerosis, hyperlipidemia, and hyperalphalipoproteinemia caused by the abnormal kinetics of CETP. WE CLAIM: 1. An anti-human CETP monoclonal antibody or a F(ab")2 or Fab" fragment from said monoclonal antibody, wherein said monoclonal antibody inhibits cholesterol ester transfer activity of human CETP and does not inhibit cholesterol ester transfer activity of rabbit CETP at a concentration of 3 µg/ml or below, and wherein said monoclonal antibody provides a lower detection limit in an immunoassay of human CETP in non-pretreated human plasma than in human plasma pretreated by detergent or heating. 2. The hybridomas # 72-1 or # 86-2 which respectively secretes monoclonal /antibody reactive to human CETP wherein said hybridomas is selected from the group consisting of the cell line deposited under the Accession number FERM BP-4944 and the cell line deposited under Accession Number FERM BP-4945 respectively. 3. A recombinant chimeric monoclonal antibody or a F(ab")2 or Fab" fragment from said monoclonal antibody, wherein said monoclonal antibodj" comprises a variable region from the monoclonal antibody, as claimed in claim 1, and a constant region from a human immunoglobulin. 4. A recombinant humanized monoclonal antibody or a F(ab")2 or Fab" fragment from / said monoclonal antibody, wherein said monoclonal antibody comprises a part of or the whole of the complementarity determining regions of the hypervariable region from the monoclonal antibody as claimed in claim 1, framework regions of the hypervariable region from a human immunoglobulin and a constant region from a human immunoglobulin. 5. An immobilized monoclonal antibody or immobilized antibody fragment -which is prepared by immobilizing the monoclonal antibody or F(ab")2 or Fab" fragment as claimed in claim 1 on an insoluble carrier. 6. The immobilized monoclonal antibody or immobilized F(ab")2 or Fab" fragment as claimed in claim 5, wherein said insoluble carrier is selected from the group consisting of a plate, a test tube, beads, a ball, a filter and a membrane. 7. The immobilized monoclonal antibody or immobilized F(ab")2 or Fab" fragment as claimed in claim 5, wherein said insoluble carrier is one used for affinity purification. 8. A labeled monoclonal antibody or labeled antibody fragment which is prepared by labeling the monoclonal antibody or F(ab")2 or Fab" fragment as claimed in claim 1 with a labeling substance that provides a detectable signal independently or by reaction with another substance. 9. The labeled monoclonal antibody or labeled F(ab")2 or Fab" fragment as claimed in claim 8, wherein said labeling substance is selected from the group consisting of an enzyme, a fluorescent material, a chemiluminescent material, biotin, avidin and a radioisotope. 10. A kit for immunoassay to detect human CETP comprising the monoclonal antibody or F(ab")2 or Fab" fragment, as claimed in claim 1. 11. A kit for immunoassay to detect human CETP comprising the labeled monoclonal antibody or labeled F(ab")2 or Fab" as claimed in claim 8. 12. A kit for separation or purification of human CETP comprising the immobilized monoclonal antibody or immobilized F(ab")2 or Fab" fragment as claimed in claim 7. 13. A pharmaceutical composition comprising the monoclonal antibody or F(ab")2 or Fab" fragment as claimed in claim 1, and a pharmaceutically acceptable carrier for prevention and/or treatment of hyperlipidemia. 14. A composition comprising the chimeric monoclonal antibody or F(ab)2 or Fab" fragment as claimed in claim 3, and a pharmaceutically acceptable carrier. 15. A composition comprising the humanized monoclonal antibody or F(ab")2 or Fab" fragment as claimed in claim 4, and a pharmaceutically acceptable carrier. Monoclonal antibodies which has binding specificity to the human CETP (CETP inhibition activity) available as reagents for the human CETP purification or determination, and Pharmaceuticals of prevention and/or treatment for hyperlipidemia or arteriosclerosis are provided. Furthermore, the purification or the determination methods of the human CETP by using the monoclonal antibody is also provided. |
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650-CAL-2000-(13-01-2012)-FORM-27.pdf
650-CAL-2000-CORRESPONDENCE 1.1.pdf
650-CAL-2000-CORRESPONDENCE.pdf
650-cal-2000-granted-abstract.pdf
650-cal-2000-granted-claims.pdf
650-cal-2000-granted-correspondence.pdf
650-cal-2000-granted-description (complete).pdf
650-cal-2000-granted-drawings.pdf
650-cal-2000-granted-examination report.pdf
650-cal-2000-granted-form 1.pdf
650-cal-2000-granted-form 18.pdf
650-cal-2000-granted-form 2.pdf
650-cal-2000-granted-form 3.pdf
650-cal-2000-granted-form 5.pdf
650-cal-2000-granted-letter patent.pdf
650-cal-2000-granted-priority document.pdf
650-cal-2000-granted-reply to examination report.pdf
650-cal-2000-granted-specification.pdf
650-cal-2000-granted-translated copy of priority document.pdf
| Patent Number | 214665 | ||||||||||||
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| Indian Patent Application Number | 650/CAL/2000 | ||||||||||||
| PG Journal Number | 07/2008 | ||||||||||||
| Publication Date | 15-Feb-2008 | ||||||||||||
| Grant Date | 15-Feb-2008 | ||||||||||||
| Date of Filing | 22-Nov-2000 | ||||||||||||
| Name of Patentee | JAPAN TOBACCO INC. | ||||||||||||
| Applicant Address | 2-1, TORANOMON 2-CHOME, MINTO-KU, TOKYO 105 | ||||||||||||
Inventors:
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| PCT International Classification Number | C07K 16/00 | ||||||||||||
| PCT International Application Number | N/A | ||||||||||||
| PCT International Filing date | |||||||||||||
PCT Conventions:
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