论文部分内容阅读
应用密度梯度超离心结合离子交换层析分离提纯人血清载脂蛋白A－1（ApoA-I），经免疫扩散，SDS－PAGE和紫外扫描鉴定，证实其达到电泳纯．以此免疫BALB/c小鼠，应用杂交瘤技术建立了7株分泌抗人ApoA-I的杂交瘤细胞株，并对其免疫学性质进行了研究。上清效价为10－2～10－4，腹水效价为10－6～10－8．除1种MCAb为lgG2a外．其余均为IgG1。经ELISA间接法检测，证明7种McAb均只与ApoA-I发生强阳性反应．而不与人ApoA-Ⅱ、ApoB100、白蛋白、球蛋白、转铁蛋白和纤雏连接蛋白等血浆蛋白反应．免疫印迹试验证实，7种McAb在包含载脂蛋白在内的众多血浆蛋白成分中，仅持异地识别MW为28KD的ApoA-I。另外还对其中3种MCAb的相对亲和力及抗原结合位点进行了研究。
Human serum apolipoprotein A-1 (ApoA-I) was isolated and purified by density gradient ultracentrifugation coupled with ion exchange chromatography. Immunoprecipitation, SDS-PAGE and UV scanning were used to confirm the purity of apolipoprotein A-1. As a result, 7 BALB / c mice were immunized with the hybridoma technology, and seven hybridoma cell lines secreting anti-human ApoA-I were established. The immunological properties were also studied. Supernatant titer of 10-2 ~ 10-4, ascites titer of 10-6 ~ 10-8. In addition to a MCAb lgG2a outside. The rest are IgG1. By indirect ELISA detection, all seven McAbs showed strong positive reaction with ApoA-I. But not with human ApoA-Ⅱ, ApoB100, albumin, globulin, transferrin and fibronectin and other plasma proteins reaction. Immunoblotting assays confirmed that among the seven McAbs, among the numerous plasma protein components including apolipoprotein, only ApoA-I with a MW of 28 kD was exclusively identified. In addition, the relative affinity and antigen binding sites of three kinds of MCAb were also studied.