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.

中文翻译：

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快速筛选具有衣壳基因变异的 GII.17 诺如病毒株的抗原特征

引起公众兴趣的新型 GII.17 Kawasaki 2014 诺如病毒变种的出现，已取代 GII.4 成为 2014-2015 年间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。在这项研究中，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​​在基于细胞的系统中表达的纯蛋白质进行比较。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。涉及一组突变无细胞蛋白的 EIA 结果表明，Q298 是循环 1 中的关键残基。这些数据突出了新型 GII.17 的线性抗体结合特征中的基本残基。此外，它支持 CFPS 作为通过诺如病毒 P 蛋白的可扩展表达快速筛选突变体的有前途的工具。