磺胺二甲嘧啶单克隆抗体的制备及特性鉴定

Preparation and characteristic identification of monoclonal antibodies directed against sulfamethazine

采用重氮化法合成磺胺二甲嘧啶(SM_2)-人血清白蛋白(HSA)免疫抗原和SM_2^-卵清白蛋白(OVA)包被抗原。经紫外光谱扫描法确认SM_2与载体蛋白偶联成功；经计算SM_2与HSA、OVA的结合比分别为9：1和15：1。利用杂交瘤技术和有限稀释法经过5次亚克隆,得到三株特异性稳定分泌SM_2抗体的杂交瘤细胞,经鉴定该单克隆抗体免疫球蛋白亚类为IgG_1,为入链,分子量为162Ku,染色体数目90条左右,亲和常数为6．1×10^(12)M^(-1)。与其他四种磺胺药和两种载体蛋白HSA、OVA均无交叉反应。

Two artificial antigens were synthesized successfully by diazotizing method, sulfamethazine（SM2）-human serum albumin （HSA） was used for the immunogen,and SM2-ovalbumin （OVA） was used for the coating antigen. The coupled reaction was successful by confirmation of the ultraviolet scanning spectrometer,and the conjugation ratio of SM2 with HSA and OVA was 9：1 and 15：1, respectively. Using cell fusion and limiting dilution method to reclone five times to get three hybridoma strains, which could stable secret monoclonal antibody（McAb）, named CB7, BC4 and BB,2. The subtype of BC4 McAb was IgG1 and λ chain. The molecular weight was 162 Ku. The chromosomal numbers were about 90. The affinity constant was 6.1×10^12M^-1. No cross- reactivity was seen between the McAb and the other four sulfonamides, as well as the two carrier proteins.