抗胆固醇酯转运蛋白单克隆抗体的制备及其应用

Preparation of monoclonal antibodies to cholesterol ester transfer protein and its aplication in determining serum concentration

目的 自行制备胆固醇酯转运蛋白 (CETP)单克隆抗体 (McAb) ,建立人血清CETP的酶联免疫吸附测定法 (ELISA) ,并调查中国人群CETP参考值。方法 应用杂交瘤技术制备CETPMcAb。用棋盘试验确定ELISA法的反应条件 ;进行灵敏度、精密度、回收率、特异性试验。结果 制备出一株特异性好、反应性强的CETPMcAb。筛选出ELISA法最佳反应条件 ,样品稀释2 0 0 0倍 ,可检测 0 2 8～ 4 5 2mg/L的血清CETP水平 ;平均批内CV 4 9% ,平均批间CV 10 2 % ;平均回收率为 92 5 %～ 97 2 % ;特异性高。检测结果同国外试验室ELISA法高度相关。 112 8例参考人群血清CETP浓度为 1 84± 1 5 5mg/L ,女性高于男性 ,随着年龄增高 ,CETP浓度降低。冠心病患者CETP浓度为 2 66± 2 0 8mg/L ,较相应对照组显著升高 (P <0 0 1)。结论 该法具有方便、快速、准确、特异等优点 ,对了解CETP在动脉粥样硬化发生中的作用有着十分重要的意义。

Objective\ To prepare monoclonal antibodies to cholesterol ester transfer protein(CETP)and develop an enzyme linked immunosorbent assay(ELISA)for measurement of human serum CETP Methods \ The monoclonal antibodies were prepared by hybridoma techology The reaction conditions with chessboard test and the assay were established Its sensitivity,precision,recovery and specificity were tested Results\ Specific anti CETP monoclonal antibodies were prepared and used to establish a Sandwich ELISA for CETP The quantitative range is 0 28 4 52 mg/L The average intra and inter assay coefficient of variation were 4 9% and 10 2% respectively CETP level determined by this ELISA correlated well with that by foreign laboratory Within 1128 healthy subjects,the average serum CETP concentration was 1 84±1 55 mg/L,the CETP level in female was higher than that in male Serum CETP concentration was negatively correlated with age in both of male and female The average CETP concentration in patients with coronary artery diseases(2 66±2 08 mg/L)was significantly higher than that in the control( P <0 01) Conclusion\ The ELISA have been established for CETP is simple,precise and specific,and is useful for investigation of the role of CETP in development of atherosclerosis