论文部分内容阅读
在体外培养细胞中,分泌单克隆抗体类型转换的突变细胞株的产生的频率极低。通过抗体依赖的补体介导的细胞毒性方法溶解杂交瘤细胞来富集分泌IgG的转换突变细胞。经过2次细胞溶解后可使突变细胞的频率增加900倍,使下一步的亚克隆易于进行。然后利用同株选择法和酶联免疫斑点测定(ELISPOT)分离和鉴定IgG分泌细胞。酶联免疫斑点测定可检测单个抗体分泌细胞,大大提高了测定的灵敏度。大多数情况,筛选出的杂交瘤细胞分泌的抗体量更高,并保持其抗原结合特异性,甚至更高的亲合性。
In cultured cells in vitro, the production of mutant cell lines secreting monoclonal antibody types is extremely low. Hybridoma cells are lysed by antibody-dependent complement-mediated cytotoxicity to enrich for IgG-secreting, transformed mutant cells. After 2 cell lysates, the frequency of mutant cells can be increased 900-fold, making the next step of subcloning easy. IgG secreting cells were then isolated and identified by isozyme selection and enzyme-linked immunosorbent assay (ELISPOT). Enzyme-linked immunosorbent assay can detect single antibody-secreting cells, greatly improving the sensitivity of the assay. In most cases, the selected hybridoma cells secrete more antibodies and retain their antigen-binding specificity and even higher affinity.