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인간 혈장 아포지단백질 A-1 ( apo A-1 ) 에 대한 단일클론 항체 생산 및 단일클론 항체를 사용한 혈중 apo A-1 측정용 효소면역분석법 ( ELISAs ) 의 개발
이동익,남경수,한문희,김태웅,곽주원,윤미청,최성아 한국지질학회 1994 韓國脂質學會誌 Vol.4 No.1
Recently, apolipoprotein(apo) A-I has received considerable clinical attention as a better marker of inverse correlation with the presence of coronary heart disease than the customary lipid marker, high-density lipoprotein(HDL)-cholesterol. Here we report the production of marine monoclonal antibodies(Mabs) designated as MabA 12 and MabA34 against human apo A-I, recognizing the HDL in plasma. From isotyping analyses, it was found that MabA 12 and MabA34 had IgG2b and IgGI isotype, respectively. Both of the Mabs had κ light chains. The binding specificity of two Mabs was analyzed by immunoblotting and enzyme-linked immunosorbent assay(ELISA). MabA12 and MabA34 reacted well with purified apo A-I or HDL in plasma but they didn't react with low-density lipoproteins(LDL), serum albumin or lipoprotein-deficient serum(LPDS). These indicate that the two Mabs are very specific to human apo A-I in HDL. The validity of the Mabs for serum apo A-I assays was examined by two kinds of ELISA, i.e., non-competitive and competitive ELISA. Both assays revealed that MabA12 or MabA34 reacted well with apo A-I in sample in concentration-dependent manners. Standard curves and, antibody titration curves were drawn. In conclusion, two IgG-type monoclonal antibodies specific to human apo A-I in HDL were produced and applicated for ELISAs of serum apo A-I. The two Mabs would be useful not only for research purposes but also for further development of immuno-diagnostic kits for the measurement of serum apo A-I concentrations.
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