抗人CD38单克隆抗体的制备、鉴定及其功能的初步研究

目的：研制抗人CD38抗原分子的单克隆抗体,进一步研究其生物学功能。方法：采用高表达CD38抗原的Daudi细胞免疫Balb/c小鼠;取其脾细胞与SP2/0细胞融合;用间接免疫荧光法进行杂交瘤筛选;流式细胞术、免疫沉淀法与CD38分子原核表达产物的Western-blot分析鉴定单克隆抗体的特异性。以MTT（四甲偶氨唑盐）法检测单克隆抗体抑制细胞增殖以及介导补体杀伤靶细胞的功能。结果：获得了一株抗人CD38分子的单克隆抗体1C6,流式细胞术显示它具有CD38抗原特异的细胞反应谱,其识别的抗原分子质量为45000u。与CD38分子原核表达产物的Western-blot分析表明,其可特异识别CD38分子胞外结构域。MTT法显示其可介导补体杀伤靶细胞。结论：成功制备了抗人CD38分子的单克隆抗体,并进行了初步的功能研究。

Preparation, characterization and function assay of monoclonal antibodies to human CD38

ObjectiveTo prepare and characterize mAb to human CD38 , and further, to study the biological function of this molecule Methods Daudi cell line which highly expresses CD38 molecule was used as antigen to immunize Balb/c mice.The spleen cells of the immunized mice were used to pr epare the McAb by hybridoma techniques.Hybridoma cells was selected by indirec t immunofluorescence experiments, immunoprecipitation and flow cytometry (FCM) a nalysis and western blot analysis with the recombinant CD38 protein were used t o identify specificity of the prepared McAb. CDC experiment and cell proliferation inhibition experiment w ere examined by MTT ResultsA monoclonal antibody against CD38 was obta ined by hybridoma techniques and named as 1C6. The results of immunoprecipitati on showed that the antibody could recognize 45 000 u molecules on the surface of Daudi cell. FCM analysis indicated that 1C6 accorded with anti CD38 McAb in s p ecific reaction with cell lines. Western blot analysis revealed that 1C6 could r ecognize the recombinant CD38 protein. Functional assay revealed that 1C6 could markedly kill target cells by CDC(complement dependant cytotoxicity).Co nclusion A hybridoma cell line 1C6 secreting anti CD38 McAb stably wa s established and characterized.