摘要
从人胎盘提取的Ⅳ型胶原(COLⅣ)为抗原,免疫BALB/c小鼠,取脾细胞与同系鼠源性的SP2/0骨髓瘤细胞,在PEG 4 000介导下融合。经克隆、筛选获得3个分泌抗COL Ⅳ单克隆抗体的杂交瘤细胞株,间接ELIsA测定表明3株抗体与COL Ⅳ有较强的抗原抗体反应,与其它3种胶原及4种测试蛋白无交叉反应。其中2株亲和力都在1011以上,此单抗可用于建立胶原Ⅳ检测方法,为临床应用提供了基础。
Type Ⅳ collagen extracted from human placenta was used as the antigen to immunize BALB/c mice. The splenocytes were fused with SP2/0 myeloma cells of synlogous mice by PEG 4 000. Three hybridoma cell lines secreting anti-type Ⅳ collagen monoclonal antibodies were gained by clonal selection . Indirect ELISA indicated that they were all extensively immuno-reactive to type Ⅳ collagen, but not cross-reactive to the other kinds of collagens or to the four kinds of test proteins. Besides, two lines of the monoclonal anti-bodies have a high affinity, over 1011. The results showed that the monoclonal anti-bodies can be used to establish an assay of collagen Ⅳ and provide a foundation for clinical application.
