论文部分内容阅读
目的：制备人亲环素Ａ特异性单克隆抗体并鉴定其特性。方法：利用基因重组技术大量表达人亲环素Ａ蛋白，纯化后作为抗原免疫ＢＡＢＬ／ｃ小鼠，经细胞融合和筛选，制备特异单克隆抗体。结果和结论：共建立三株稳定分泌抗人ＣｙＰＡ的杂交瘤细胞株，其所产生的单克隆抗体均为ＩｇＭ型，轻链为κ。ＥＬＩＳＡ和点印迹试验表明，三株ＭｃＡｂ均与人ＣｙＰＡ标准品反应，免疫印迹试验证明只与大肠杆菌中分子量１８０００重组人ＣｙＰＡ组份反应。
OBJECTIVE: To prepare human cyclophilin A specific monoclonal antibody and identify its characteristics. Methods: The gene of human cyclophilin A was expressed in large quantities by gene recombination technique. After being purified, it was used as antigen to immunize BABL / c mice. The specific monoclonal antibodies were prepared by cell fusion and screening. RESULTS AND CONCLUSION: Three hybridoma cell lines stably secreting anti-human CyPA were established. All the monoclonal antibodies produced were of IgM type and the light chain was κ. ELISA and dot blot assays showed that all three McAbs reacted with human CyPA standards and Western blotting assays demonstrated only reaction with a molecular weight of 18,000 recombinant human CyPA components in E. coli.