Department of Food Science and Technology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.
State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangdong Institute of Microbiology, Guangzhou, 510070, China.
Virol Sin. 2020 Feb;35(1):103-109. doi: 10.1007/s12250-019-00174-5. Epub 2019 Nov 27.
Rotavirus (RV) is a major foodborne pathogen. For RV prevention and control, it is a key to uncover the interaction mechanism between virus and its receptors. However, it is hard to specially purify the viral receptors, including histo-blood group antigens (HBGAs). Previously, the protruding domain protein (P protein) of human norovirus (genotype II.4) was displayed on the surface of Escherichia coli, and it specifically recognized and captured the viral ligands. In order to further verify the feasibility of the system, P protein was replaced by VP8* of RV (G9P[8]) in this study. In the system, VP8* could be correctly released by thrombin treatment with antigenicity retaining, which was confirmed by Western blot and Enzyme-Linked Immunosorbent Assays. Type A HBGAs from porcine gastric mucin (PGM) were recognized and captured by this system. From saliva mixture, the captured viral receptor bound with displayed VP8* was confirmed positive with monoclonal antibody against type A HBGAs. It indicated that the target ligands could be easily separated from the complex matrix. These results demonstrate that the bacterial surface display system will be an effective platform to explore viral receptors/ligands from cell lines or food matrix.
轮状病毒(RV)是一种主要的食源性病原体。为了预防和控制 RV,揭示病毒与其受体的相互作用机制是关键。然而,专门纯化病毒受体(包括组织血型抗原(HBGAs))非常困难。以前,人诺如病毒(基因型 II.4)的突出结构域蛋白(P 蛋白)在大肠杆菌表面展示,并特异性识别和捕获病毒配体。为了进一步验证该系统的可行性,本研究用 RV(G9P[8])的 VP8替代 P 蛋白。在该系统中,VP8可通过凝血酶处理正确释放,且保留抗原性,这通过 Western blot 和酶联免疫吸附试验得到证实。来自猪胃粘蛋白的 A 型 HBGAs 被该系统识别和捕获。从唾液混合物中,与展示的 VP8*结合的捕获的病毒受体用针对 A 型 HBGAs 的单克隆抗体被证实为阳性。这表明目标配体可以很容易地从复杂基质中分离出来。这些结果表明,细菌表面展示系统将成为从细胞系或食物基质中探索病毒受体/配体的有效平台。