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目的:制备抗人层黏连蛋白(laminin,LN)单克隆抗体(mAb)并鉴定其特性。方法:以人LN免疫BALB/c小鼠,采用杂交瘤技术制备抗人的mAb;同时采用间接ELISA法柃测mAb的腹水效价及mAb的相对亲和力;采用ELISA法鉴定mAb的Ig亚类、进行表位分析及特异性鉴定。结果:获得4株可分泌特异性mAb的杂交瘤细胞2A3、2C6、3G7和4H2,其腹水mAb 的效价为3.6 × 104～2.1×106;4株mAb的Ig亚类为IgG1,轻链均属κ型;相对亲和力2C6在1012以上,2A3、3G7和4H2在106以上;其中2株与1个表位结合,另2株与另外的1个表位结合。结论:成功地制备出抗人LN的mAb,为进一步研究LN在一些疾病中的作用提供了工具。
Objective: To prepare monoclonal antibodies against human laminin (LN) and to identify its characteristics. Methods: BALB / c mice were immunized with LN and anti-human mAb was prepared by hybridoma technique. The indirect ELISA was used to measure the ascites potency of mAb and the relative affinity of mAb. The Ig subclass of mAb was identified by ELISA. For epitope analysis and specific identification. Results: Four hybridoma cells secreting specific mAbs 2A3, 2C6, 3G7 and 4H2 were obtained. The titer of ascites mAb was 3.6 × 104 ~ 2.1 × 106. The Ig subclass of 4 mAbs was IgG1, Is a κ-type; the relative affinity of 2C6 in more than 1012, 2A3,3G7 and 4H2 in more than 106; of which two strains with one epitope, the other two with another epitope binding. Conclusion: The successful preparation of anti-human LN mAb provides a tool to further study the role of LN in some diseases.