抗戊型肝炎病毒重组蛋白单克隆抗体的制备和鉴定

Generation and characterization of monoclonal antibodies against ORF23 recombinant protein of hepatitis E virus

目的:制备能够特异性识别戊型肝炎病毒重组蛋白ORF23的单克隆抗体并分析其生物学特性。方法:利用纯化的戊型肝炎病毒(HEV)ORF23基因工程重组蛋白免疫BALB/c小鼠,采用常规杂交瘤细胞技术制备单克隆抗体,并对其单克隆抗体的相关性质进行鉴定。结果:获得两株稳定分泌抗体的杂交瘤细胞株A8和A11。间接ELISA方法测定,杂交瘤细胞培养上清效价均为1:32000,接种小鼠的腹水效价均为1:2×106。两株单抗均能特异性识别重组ORF23蛋白,但它们识别ORF23蛋白的不同表位。结论:获得的单克隆抗体可以为诊断HEV提供敏感、特异、稳定的诊断试剂,并为HEV ORF23重组蛋白抗原表位的精确定位以及基于表位的基因工程疫苗研究奠定基础。

Objective：To generate and characterize the McAbs against ORF23 recombinant protein of HEV. Methods：Five - weeks - old BALB/c mice were immunized with purified recombinant protein ORF23 of hepatitis E virus. The immunized spleen cells were fused with SP2/0 to raise hybridoma. Results： Two hybridoma cell lines designated A8 and All steadily secreting McAbs against ORF23 protein of hepatitis E virus were obtained by hybridoma technology. The titers of the two McAbs in the culture supernatant were 1：32000, respectively, and the titers in the ascites of the mice were all 1：2 × 10^6 detected by ELISA. The McAbs were able to recognize different epitopes of ORF23 and have the specificity. Conclusion ： The McAbs can be used to develop the HEV diagnostic kit, and also analyze the antipopes of HEV.