3α-HSD单克隆抗体的制备及其鉴定

Development and Identification of Monoclonal Antibody Against 3α-HSD

为制备抗3α-HSD蛋白单克隆抗体(MAb),以已表达纯化的高纯度3α-HSD蛋白作为免疫原免疫BALB/c小鼠,利用杂交瘤技术,经过免疫和细胞融合后筛选出特异性的抗3α-HSD蛋白单克隆抗体,成功冻存35株IgG类型阳性杂交瘤细胞株。对14号和20号Anti-3α-HSD进行亲和性测试,发现其与可溶性抗原蛋白的亲和常数分别为3.7E+09和4.2E+09,二者可与抗原牢固结合,具有良好的亲和性。

For development of monoclonal antibody（MAbs） against 3α-HSD, BALB/c mice were immunized with a synthesized 3α- HSD. The splenocytes of the immunized mice were fused with sp2/0 cells,and the positive hybridoma cells were screened. 35 of MAbs were identified as IgG and successfully cryopresevated, the affinity constant of 14# and 20# MAbs against 3α-HSD were 3.7E＋09 and 4.2E＋09.The affinity of MAbs was good.