中国动物传染病学报2024，Vol.32Issue(3)：19-26,8.
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
摘要
Abstract
The synthetic peptide antibodies of PRRSV non-structural protein Nsp1α have not been reported yet.In this study,the synthetic peptide of PRRSV Nsp1α was used to prepare polyclonal antibodies for further analysis.First,the antigenic peptide sequence of PRRSV non-structural protein Nsp1α was 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 Nsp1α band 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 Nsp1α could be used in the detection of PRRSV Nsp1α expression,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Key words
Porcine reproductive and respiratory syndrome virus/synthetic peptide/polyclonal antibody/nonstructural protein分类
农业科技引用本文复制引用
解艺璇,李蓓蓓,马志永,孙延鸣,邱亚峰,张彦兵,李慧,朱琳,葛菲菲,刘珂,魏建超,李宗杰,邵东华..PRRSV非结构蛋白Nsp1α合成肽多克隆抗体的制备及应用[J].中国动物传染病学报,2024,32(3):19-26,8.基金项目
上海市自然科学基金(22ZR1475900) （22ZR1475900）
中央级公益性科研院所基本科研业务费(2020JB04) （2020JB04）
上海市科技兴农项目(沪农科推字(2020)第1-4号) （沪农科推字(2020）
