PRRSV非结构蛋白Nsp1α合成肽多克隆抗体的制备及应用  

Characterization of Polyclonal Antibodies Against PRRSV Nonstructural Protein 1α(Nsp1α)Generated by Immunization with a Synthetic Peptide

作　　者：解艺璇 张彦兵[1] 李慧 朱琳 葛菲菲[3] 刘珂[2] 魏建超[2] 李宗杰 邵东华[2] 李蓓蓓[2] 马志永[2] 孙延鸣[1] 邱亚峰[2] XIE Yixuan;ZHANG Yanbing;LI Hui;ZHU Lin;GE Feifei;LIU Ke;WEI Jianchao;LI Zongjie;SHAO Donghua;LI Beibei;MA Zhiyong;SUN Yanming;QIU Yafeng(College of Animal Science and Technology,Shihezi University,Shihezi 832003,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China;Shanghai Animal Disease Prevention and Control Center,Shanghai 201103,China)

机构地区：[1]石河子大学动物科技学院,石河子832003 [2]中国农业科学院上海兽医研究所,上海200241 [3]上海市动物疫病预防控制中心,上海201103

出　　处：《中国动物传染病学报》2024年第3期19-26,共8页Chinese Journal of Animal Infectious Diseases

基　　金：上海市自然科学基金(22ZR1475900);中央级公益性科研院所基本科研业务费(2020JB04);上海市科技兴农项目(沪农科推字(2020)第1-4号)。

摘　　要：本研究尝试利用PRRSV Nsp1α合成肽进行多克隆抗体的制备,并对抗体的特性进行了分析。首先,利用生物信息学技术预测获得PRRSV非结构蛋白Nsp1α的抗原肽序列;随后,按照此序列合成获得抗原肽,并将其与KLH载体蛋白进行偶联;接下来,将偶联好的抗原肽与弗氏佐剂混合,通过免疫大白兔制备针对Nsp1α合成肽的多克隆抗体;最后,利用ELISA、Western blot及间接免疫荧光法对多克隆抗体的特性进行了分析。结果显示:该多克隆抗体ELISA的效价超过105;利用Western blot可以检测到特异的Nsp1α表达条带,包括瞬时转染的样品和不同毒株病毒感染的样品;利用间接免疫荧光法分析,该多克隆抗体可有效地区分病毒感染和未感染的样品。结果表明,PRRSV Nsp1α合成肽多克隆抗体可以有效地应用于PRRSV Nsp1α表达检测,为进一步研究PRRSV Nsp1α的功能提供了一个有效的工具。The synthetic peptide antibodies of PRRSV non-structural protein Nsp1αhave not been reported yet.In this study,the synthetic peptide of PRRSV Nsp1αwas used to prepare polyclonal antibodies for further analysis.First,the antigenic peptide sequence of PRRSV non-structural protein Nsp1αwas predicted by using bioinformatics technology.Then,the antigenic peptide was synthesized according to this sequence and coupled with the KLH carrier protein.The coupled antigenic peptide was emulsified with Freund's adjuvant to immunize rabbits for preparation of polyclonal antibodies.Finally,the polyclonal antibodies were analyzed in ELISA,Western blot and indirect immunofluorescence.The results show that the ELISA titer of the polyclonal antibodies exceeded 105 and the specific Nsp1αband was visualized in Western blotting,including samples from transient transfection and infection with different virus strains.Additionally,the polyclonal antibodies confirmed virus-infected and uninfected samples in indirect immunofluorescence.Therefore,the synthetic peptide polyclonal antibodies against PRRSV Nsp1αcould be used in the detection of PRRSV Nsp1αexpression,thus providing an effective tool for further research on the function of PRRSV Nsp1α.

关 键 词：猪繁殖与呼吸综合征病毒 合成肽 多克隆抗体 非结构蛋白 

分 类 号：S859.797[农业科学—临床兽医学]