猫杯状病毒单克隆抗体的制备
Development of Monoclonal Antibodies against Feline Calicivirus
王莹 1李佳康 2曾悦 1史开拓 1韦丽婷 1彭佳佳 1李秋燕 1曹龙龙 2周登元3
作者信息
- 1. 武汉科前生物股份有限公司,武汉 430000
- 2. 华中农业大学动物科学技术学院动物医学院,武汉 430070
- 3. 武汉科前生物股份有限公司,武汉 430000;华中农业大学动物科学技术学院动物医学院,武汉 430070
- 折叠
摘要
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3B110 MAb 3B11抗体类型鉴定其重链属于IgG1、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.
Abstract
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 and κ chain 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.
关键词
猫杯状病毒/单克隆抗体/特异性/反应性Key words
feline calicivirus/monoclonal antibodies/specificity/reactivity引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据