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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 prepare monoclonal antibodies that can specifically recognize ORF23 of hepatitis E virus recombinant protein and analyze its biological characteristics. Methods: BALB / c mice were immunized with recombinant HEV ORF23 gene recombinant protein. The monoclonal antibodies were prepared by conventional hybridoma technique and the related properties of the monoclonal antibodies were identified. Results: Two hybridoma cell lines stably secreting antibodies, A8 and A11, were obtained. Indirect ELISA assay, the hybridoma cell culture supernatant titer was 1: 32000, inoculation mice ascites titer were 1: 2 × 106. Both monoclonal antibodies specifically recognize the recombinant ORF23 protein, but they recognize different epitopes of the ORF23 protein. Conclusion: The obtained monoclonal antibodies can provide sensitive, specific and stable diagnostic reagents for HEV diagnosis and lay a foundation for the precise localization of HEV ORF23 recombinant protein epitopes and epitope-based genetic engineering vaccine research.