β-内酰胺酶单克隆抗体和多克隆抗体的制备

Preparation of Monoclonal Antibody and Polyclonal Antibody Against β-Lactamase

比较β-内酰胺酶和青霉素酶两种蛋白的纯度,选择β-内酰胺酶为免疫原。通过免疫家兔制备抗β-内酰胺酶多克隆抗体,4次免疫后,抗体的效价均在1×106以上。同时免疫Balb/C小鼠,取免疫后的脾细胞与SP2/0细胞进行化学融合,使用青霉素酶筛选阳性杂交瘤细胞株;4次亚克隆后将阳性细胞株腹腔注射BALB/C小鼠,获得阳性腹水;用PEG6000法纯化腹水,制备单克隆抗体;对该单克隆抗体进行特性分析,为制备检测β-内酰胺酶的双抗体夹心胶体金免疫层析试纸条奠定基础。

Compared of the purity between the β-Lactamase and the penicillinase, the β-1actamase was selected as the immunogen. The Anti-β-1actamase polyelonal antibody was prepared by immunizing rabbits. Working concentration of rabbit anti-seras were upon 1×10^6. At the same time the BALB/C mice were immuned, the splenocytes of the immunized mice were fused with SP2/0 cells, the positive hybridoma cells were screened by the penicillinase. After the four sub- clones, the positive cell lines was injected into the abdominal cavity of BALB/C mice to obtain positive ascites. The as- cites were purified by PEG6000 for the preparation of monoclonal antibodies. The monoclonal antibodies were conformed. The experiment laid the foudation for the study of Colloidal gold immunochromatography test strip by Double antibody sandwich method.