抗HCV HVR1模拟合成肽单克隆抗体的制备及特性鉴定  

Preparation and characterization of the anti-HCV HVR1 synthetic peptide monoclonal antibody

作　　者：孙杰[1] 戚扬[1] 迟淑萍[1] 由莉越[1] 李瑞生[1] 刘佳[1] 朱雷[1] 程云[1] 

机构地区：[1]解放军302医院传染病研究所免疫室,北京100039

出　　处：《细胞与分子免疫学杂志》2011年第5期542-544,共3页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的:制备抗-HCV高变区1(HVR1)合成肽单克隆抗体(mAb),并对其特性进行鉴定。方法:以固相合成的HCV HVR1合成肽与牛血清白蛋白(BSA)偶联后,免疫8周龄的BALB/c小鼠。采用杂交瘤技术,取免疫小鼠的脾细胞与骨髓瘤细胞融合,制备抗-HCV HVR1 mAb。经HAT、HT选择培养及有限稀释法进行克隆化后,用相关的试剂盒鉴定小鼠mAb的Ig亚类;经中和抑制后用ELISA法检测mAb的特异性;用间接ELISA法检测mAb腹水的效价及相对亲和常数。结果:获得1株可分泌抗-HCV HVR1合成肽特异性mAb的杂交瘤细胞1A9G9F11。该株mAb的Ig亚类为IgG3;腹水效价为3.125×10-5;相对亲和常数为1.0×106L/mol。该株mAb与HBsAg、HBeAg、HBcAg、HAAg、牛血清白蛋白、酪蛋白和胸腺5肽均无交叉反应,与HCV HVR1合成肽-牛血清白蛋白出现特异性反应。结论:成功地建立了1株可分泌抗-HCV HVR1 mAb的杂交瘤细胞1A9G9F11,为进一步的实验研究提供了重要的制剂。AIM： To prepare monoclonal antibody（mAb） against HCV HVR1 synthetic peptide and characterize its property.METHODS： 8 week-old female BALB/c mice were immunized with HCV HVR1 synthetic peptide which was linked to bovine serum albumin（BSA） as immunogen.The splenocytes of immunized mice were fused with Sp2/0 cells by hybridoma technique.Monoclonal hybridoma cell line was obtained by limiting dilution,HAT and HT-selective culture.The specificity of mAb was identified by indirect ELISA,immunohistochemical staining and inhibitory reactivity.RESULTS： One hybridoma cell（1A9G9F11） was obtained.The mAb belonged to Ig G3.The titers of mAb in ascetic and the relative affinity constant of mAb were 3.125×10-5 and 1.0×106 L/mol respectively.Indirect ELISA detecition demonstrated that the mAb had no cross-reactivates with other viral antigens,such as HBsAg,HBeAg and HAAg,and with other protein,such as BSA,casein and thymopentin.CONCLUSION： One hybridoma cell secreting mAb against-HCV HVR1 has been established successfully,which lays the foundationfor for the further study.

关 键 词：HCVHVR1 合成肽 单克隆抗体 特性鉴定 

分 类 号：R392.11[医药卫生—免疫学]