非洲猪瘟病毒p54蛋白特定抗原表位单克隆抗体的制备

Preparation of monoclonal antibody against specific antigenic epitope of the p54 protein of African swine fever virus

为了制备针对非洲猪瘟病毒p54蛋白已知特定抗原表位的单克隆抗体,采用原核表达的重组蛋白p54免疫小鼠,并制备了原核表达p54蛋白、真核表达p54蛋白以及人工合成特定抗原表位氨基酸多肽A三种筛选抗原,筛选出了1株针对已知抗原位点的杂交瘤细胞株。对其分泌的单克隆抗体进行了亚类及特异性鉴定,并利用ProteOn XPR36大分子相互作用系统分析了该单克隆抗体的亲和性。结果显示,该单克隆抗体属IgG1亚类,特异性强、亲和力高。这为以后的免疫学检测方法的建立奠定了基础。

In order to prepare monoclonal antibody(McAb)against known specific antigenic epitope of the p54 protein of African swine fever virus,mice were immunized with the recombinant protein p54 that is expressed in prokaryotic system.Antigens used for screening the prepared McAb include the p54 protein of prokaryotic expression,the p54 protein of eukaryotic expression protein and the synthetic specific antigenic epitope polypeptide of p54.A hybridoma secreting McAb against the specific antigenic epitope of p54 was generated by fusion of the SP2/0cells with the splenic cells from the recombinant protein p54 immunized BALB/c mice.Here was identified characteristic of McAb,and the ProteOn XPR36 interaction array system was used to analyze McAb affinity.In result,the monoclonal antibody belongs to IgG1 subspecies,and it shows a good specificity and affinity.The result laid the foundation for the establishment of immunological detection method.