猫杯状病毒单克隆抗体的制备

Development of Monoclonal Antibodies against Feline Calicivirus

制备猫杯状病毒(feline calicivirus,FCV)的特异性单克隆抗体,为FCV的免疫学诊断方法提供材料。本研究将经蔗糖梯度离心纯化的FCV灭活后免疫BALB/c小鼠,通过间接ELISA方法筛选阳性杂交瘤细胞,并对其进行一系列鉴定。结果成功获得1株能够稳定分泌抗体的单克隆细胞株3B11。MAb 3B11抗体类型鉴定其重链属于IgG_(1)、轻链为κ链。MAb 3B11能与FCV-SH192发生特异性反应;小鼠腹水MAb 3B11对5株FCV毒株具有较高的ELISA效价;与5株FCV毒株具有间接免疫荧光(IFA)和蛋白免疫印迹(Western blot,WB)结合特性。本研究制备的MAb 3B11对FCV具有良好的特异性、免疫反应性及广谱识别性,为FCV诊断方法的优化、流行毒株筛选以及基础研究提供生物原料。

This study aimed to develop specific monoclonal antibodies for feline calicivirus(FCV)and provide materials for immunological diagnostic methods of FCV.In this study,BALB/c mice were immunized with FCV purified by sucrose gradient centrifugation and inactivated.The positive hybridoma cells were screened by an indirect ELISA method and characterized by a series of methods.One hybridoma 3B11 stably secreting monoclonal antibodies was successfully obtained.The antibody type of MAb 3B11 was identified as IgG1 for the heavy chain andκchain for the light chain.MAb 3B11 specifically reacted with FCV-SH192.The mouse ascites MAb 3B11 showed high ELISA titers against five FCV strains;it had IFA and Western blot binding characteristics with five FCV strains.The MAb 3B11 developed in this study has good specificity,immunoreactivity and broad-spectrum recognition of FCV,which provides biological raw materials for the optimization of FCV diagnostic methods,screening of epidemic strains and basic research.