邻氯青霉素单克隆抗体的制备及特性鉴定

采用碳化二亚胺法，将邻氯青霉素（cloxacillin）与牛血清白蛋白偶联制备免疫原，免疫BALB/c小鼠，经杂交瘤技术获得了一株能稳定分泌抗邻氯青霉素单克隆抗体的杂交瘤细胞株2H8-B1-F4。间接ELISA方法测定，抗体亚类为IgG1，其细胞上清抗体效价为1：1000，腹水效价为1：4×105。与其结构类似物苯唑青霉素、双氯青霉素的交叉反应率为21.3％和19.6％，和氨苄青霉素、羟氨苄青霉素和苄青霉素无交叉反应。体外传代培养和冻存复苏后抗体分泌稳定。为进一步研制检测邻氯青霉素的 ELISA试剂盒奠定基础。

PREPARATION AND IDENTIFICATION OF MONOCLONAL ANTIBODY AGAINST CLOXACILLIN

CX-conjugated antigen(BSA-CX) were produced by by methods of EDC.The spleen cells of BALB/C mouse immunized by BSA-CX were fused with SP2/0 plasmacytoma cells using PEG4000,and selected cultured with HAT and HT medium.A hybridoma cell line of 2H8-B1-F4 was secreened for specificity to CX,and cloned by limited dilution method for 3 times,which secrete high affinity and stable monoclonal antibodies against CX with indirect ELISA. The antibody was characterized by IgG1 isotype, the titers of antibody in cell supernatant and ascites were 1:1000 and 1:4×105,respectively.The mAb against CX was selective toward Dicloxacillin、Oxacillin with cross-reactivity of about 19.6％ and 21.3％,which can be used to detect CX residues by competitive ELISA.