猪圆环病毒2型Rep蛋白单克隆抗体的制备及鉴定

Preparation and Identification of Monoclonal Antibodies against Rep Protein of Porcine Circovirus Type 2

本研究旨在制备猪圆环病毒2型(PCV2)Rep蛋白单克隆抗体。PQE30-PCV2Rep蛋白在IPTG的诱导下表达,用SDS-PAGE对表达产物进行检测。将纯化的重组蛋白免疫小鼠,制备单克隆抗体,ELASA法鉴定抗体亚型及效价。结果显示重组蛋白PQE30-PCV2Rep得到高效表达,筛选获得了3株稳定分泌抗Rep蛋白单克隆抗体的杂交瘤细胞系1G4、2G4和6H12,其分泌的抗体分别为IgG2a、IgG2b、IgG2b亚型。其中,6H12抗体效价为256K,浓度为0.47mg/mL,纯度>90%。本研究制备的单克隆抗体有较好的生物学特性,为PCV2的研究及快速诊断提供了新的方法。

The objective of this study is to prepare monoclonal antibodies against Rep protein of porcine circovirus type 2(PCV2).The PQE30-PCV2 Rep protein was expressed under IPTG induction,and the expression products were identified by SDS-PAGE.The purified recombinant protein was used to immunize mice in order to prepare monoclonal antibodies,and ELASA method was used to identify antibody subtypes and titers.The results showed that the recombinant protein PQE30-PCV2 Rep was highly expressed,and 3 hybridoma cell lines 1G4,2G4 and 6H12 monoclonal antibodies against Rep protein were screened out,their antibody subtypes were IgG2a,IgG2b and IgG2b,respectively.The titer of 6H12 antibody was 256K,the concentration was 0.47 mg/mL,and its purity was>90%.The monoclonal antibodies prepared in this study have good biological characteristics,which provides a new method for the research and rapid diagnosis of PCV2.