摘要
为优化口蹄疫纳米抗体M170和M8蛋白的制备工艺,通过构建原核及真核重组表达质粒,分别在大肠杆菌或悬浮HEK293细胞中表达上述2种重组抗体蛋白,并对其纯度、产量以及亲和力进行比较分析。结果表明,原核表达系统制备M170和M8重组蛋白的产量是真核表达系统的2.5~3倍,生产周期可缩短至2 d内,适合进行规模化生产;真核表达系统制备M170和M8重组蛋白至少需要7 d、且产量较低;蛋白活性鉴定结果表明,两种表达系统制备的M170和M8构象无显著差异,都能特异性识别口蹄疫病毒146S抗原或12S抗原。本研究为后期口蹄疫病毒型抗原定量ELISA试剂盒的生产工艺优化提供了参考。
order to optimize the preparation process of foot-and-mouth disease(FMD)nanoantibody M170 and M8 proteins,recombinant expression plasmids were constructed to express the two recombinant proteins by E.coli or HEK293 cells,respectively.The purity,yield and affinity of M170 and M8 were compared and analyzed.The results showed that the yield of M170 and M8 prepared by prokaryotic expression system was 2.5~3 times than that of eukaryotic expression system,and the production cycle of prokaryotic expression system could be shortened to 2 days,which was suitable for large-scale production,but the preparation of M170 and M8 by eukaryotic expression systems need at least 7 days and yields were low.the results of protein activity test showed that there was no significant difference in conformation between M170 and M8 prepared by the two expression systems,and both could specifically recognize foot-and-mouth disease virus 146S antigen or 12S antigen.This study provided a reference for the production process optimization of O-type FMD antigen quantitative ELISA kit.
作者
刘仲元
南栩
谭书桢
秦峰
董虎
张韵
郭慧琛
孙世琪
徐业芬
LIU Zhongyuan;NAN Xu;TAN Shuzhen;QIN Feng;DONG Hu;ZHANG Yun;GUO Huichen;SUN Shiqi;XU Yefen(Key Laboratory of Hydatidosis Control,Ministry of A griculture and Rural Affairs/Provincial Key Laboratory of Xizang Plateau A nimal Epidemic Disease Research/College of Animal Sciences,Xizang A griculture and Animal Husbandry College,Linzhi 860000,China;State Key Laboratory for A nimal Disease Control and Prevention/College of Veterinary Medicine of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of A gricultural Sciences,Lanzhou 730000,China)
出处
《中国兽医科学》
北大核心
2025年第7期919-924,共6页
Chinese Veterinary Science
基金
甘肃省自然科学基金项目(24JRRA012)
国家自然科学基金青年基金项目(32301127)
2023兰州市人才创业创新项目(2023-RC-3)。
关键词
口蹄疫病毒
146S抗原
纳米抗体
foot-and-mouth disease virus
146S antigen
nano-antibody
