利巴韦林免疫原的制备

Preparation of ribavirin immune antigen

【目的】通过抗原合成、免疫小鼠制备单克隆抗体,为研制利巴韦林检测试剂盒和胶体金免疫层析试纸条奠定基础。【方法】通过琥珀酸酐法制备利巴韦林的完全抗原R-HS-BSA,免疫BALB/c小鼠,使小鼠产生免疫反应。建立间接ELISA和间接竞争ELISA系统,检测小鼠血清,并应用杂交瘤技术建立利巴韦林Mab细胞株,用体内诱生腹水法制备利巴韦林单克隆抗体,并对其效价、敏感性免疫学特性进行鉴定。【结果】成功合成利巴韦林的免疫原R-HS-BSA和包被原R-HS-OVA,二者蛋白浓度为10.9mg/mL和10.6mg/mL,结合比为111和91;血清效价为132 000;制备的单克隆抗体浓度为7.462mg/L;得到单克隆抗体的IC50为39.8ng/mL。【结论】初步建立利巴韦林的完全抗原及包被原的制备方法,得到利巴韦林单克隆抗体。

[Objective]To develop a method for the rapid detection of Ribavirin（R）, Ribavirin monoclonal antibody was pre- pared. [Methods]In this study, Ribavirin complete antigen（R-HS-BSA） was synthesized using succinic anhydride method. BALB/c mouse were immunized using the complete antigen of R-HS-BSA. The mice serum was tested by the indirect ELISA and indirect competitive ELISA method. Ribavirin monoclonal antibody cell line was developed through splenocyte hybridoma technique. Ascites for ribavirin monoclonal antibody was induced in mouse abdomen, and the titer, immunological properties such as sensitivity and specificity were identified. [Results] Complete immunogenic R-HS-BSA and coating antigen R-HS-OVA were synthesized, The protein concentration were 10.9 mg/mL for R-HS-BSA and 10.6 mg/mL for R-HS-OVA, respective- ly. The coupling ratio of R to BSA was 13 ： 1 and to OVA was 9 ： 1. The titer of antiserum was 1 ： 32 000, The antibody concen- tration in serum was 7. 462 mg/mL. [Conclusion]We preliminarily synthesized the Ribavirin complete antigen, and prepared ribavirin monoclonal antibody.