禽传染性支气管炎病毒S1蛋白单克隆抗体制备及其中和抗原表位的鉴定Preparation of monoclonal antibody against S1 protein of avian infectious bronchitis virus and identification of its epitopes
彭莉,张焕东,李忠斐,肖鹏,颜焰,董伟仁,周继勇,廖敏
摘要(Abstract):
为鉴定禽传染性支气管炎病毒(IBV)刺突(S1)蛋白的中和抗原表位,本研究通过IBV全病毒免疫BALB/c小鼠,经融合、亚克隆筛选获得了4株稳定分泌抗IBV S1蛋白抗体的杂交瘤细胞株4A11、1B11、5E5和7C9。经鉴定制备的单克隆抗体腹水抗体ELISA效价为10~6以上,Western blot和间接免疫荧光试验分析显示该单克隆抗体反应性和特异性良好。随后用8种基于S1分型的IBV毒株为试验毒株,同实验室已有的8株IBV S1单抗(1E9、1H1、1E4、3C6、3C7、2F3、2E5、4F9)共12株单抗进行气管环中和活性检测,发现S1蛋白单抗与8种S1亚型的毒株均有不同程度的中和作用。随后,在抗原表位和Western blot分析的基础上,鉴定出单克隆抗体识别的抗原表位区域,获得了6个中和抗原表位,其中除~(416)IQTRTEP~(422)表位,其他5个表位仍未有相关报道。本研究成功制备出4株特异性识别S1蛋白且具有中和活性的单克隆抗体,不仅丰富了IBV的单克隆抗体库,为今后研究IBV S1蛋白分子结构提供了关键生物材料;获得的6个S1蛋白中和抗原表位也为后续建立IBV疫苗免疫效果评价方法和亚单位疫苗开发提供了工具。
关键词(KeyWords): 禽传染性支气管炎病毒;S1蛋白;单克隆抗体;中和抗原表位
基金项目(Foundation): 国家重点研发计划资助项目(2021YFD1801102)
作者(Author): 彭莉,张焕东,李忠斐,肖鹏,颜焰,董伟仁,周继勇,廖敏
DOI: 10.16303/j.cnki.1005-4545.2023.12.04
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