Liao Yingyin, Xue Liang, Gao Junshan, Zuo Yueting, Liang Yanhui, Jiang Yueting, Cai Weicheng, Yang Jiale, Zhang Jumei, Ding Yu, Chen Moutong, Wu Aiwu, Kou Xiaoxia, Wu Qingping
Guangzhou Key Laboratory for Clinical Rapid Diagnosis and Early Warning of Infectious Diseases, KingMed School of Laboratory Medicine, Guangzhou Medical University, Guangzhou, China.
Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, China.
Gut Pathog. 2022 Jul 25;14(1):31. doi: 10.1186/s13099-022-00504-1.
The emergence of the novel GII.17 Kawasaki 2014 norovirus variant raising the interest of the public, has replaced GII.4 as the predominant cause of noroviruses outbreaks in East Asia during 2014-2015. Antigenic variation of the capsid protein is considered as one of the key mechanisms of norovirus evolution. In this study, we screened a panel of GII.17 mutants. First, we produced norovirus P proteins using cell-free protein synthesis (CFPS) system, comparing the results to pure proteins expressed in a cell-based system. Next, we determined the binding capability of specific monoclonal antibody (mAb) 2D11 using a unique set of wild-type GII.17 strains. Results of the EIA involving a panel of mutant cell-free proteins indicated that Q298 was the key residue within loop 1. These data highlighted the essential residues in the linear antibody binding characteristics of novel GII.17. Furthermore, it supported the CFPS as a promising tool for rapidly screening mutants via the scalable expression of norovirus P proteins.
新型GII.17 2014年川崎诺如病毒变体的出现引起了公众的关注,在2014 - 2015年期间,它已取代GII.4成为东亚诺如病毒爆发的主要原因。衣壳蛋白的抗原变异被认为是诺如病毒进化的关键机制之一。在本研究中,我们筛选了一组GII.17突变体。首先,我们使用无细胞蛋白质合成(CFPS)系统生产诺如病毒P蛋白,并将结果与基于细胞的系统中表达的纯蛋白进行比较。接下来,我们使用一组独特的野生型GII.17菌株确定了特异性单克隆抗体(mAb)2D11的结合能力。涉及一组无细胞突变蛋白的酶免疫分析结果表明,Q298是环1内的关键残基。这些数据突出了新型GII.17线性抗体结合特性中的必需残基。此外,它支持CFPS作为一种通过可扩展表达诺如病毒P蛋白快速筛选突变体的有前途的工具。