Expression of VP7 protein of group A rotavirus G1P8 and preparation of antibody to VP7
10.3760/cma.j.cn112866-20230816-00017
- VernacularTitle:人A组轮状病毒G1P[8]基因型VP7蛋白的表达及其抗体制备
- Author:
Ying ZHAO
1
;
Yunxi SU
;
Qing ZHANG
;
Huiying LI
;
Dandi LI
;
Jinsong LI
;
Xiaoman SUN
;
Zhaojun DUAN
Author Information
1. 甘肃中医药大学公共卫生学院，兰州 730000
- Keywords:
Rotavirus;
VP7 protein;
Baculovirus expression system;
Polyclonal antibody
- From:
Chinese Journal of Experimental and Clinical Virology
2023;37(6):643-649
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To express and purify VP7 protein of group A rotavirus (RVA) G1P[8]. The VP7 polyclonal antibody was prepared and its function was evaluated.Methods:The G1 VP7 protein was expressed by baculovirus expression system and purified by affinity chromatography. Polyclonal antibody against G1 VP7 was obtained by immunizing rabbits with G1 VP7 protein. The function of the G1 VP7 polyclonal antibody was verified by Western blotting (WB), enzyme-linked immunosorbent assay (ELISA), and immunofluorescence assay.Results:The soluble G1 VP7 protein of human RVA G1P[8] was obtained using the baculovirus expression system and the VP7 protein was mainly in trimer state. The G1 VP7 polyclonal antibody was prepared and displayed relatively high binding titer to G1 VP7 protein by ELISA. The VP7 polyclonal antibodies could recognize multiple G-type RVAs by WB and ELISA. Immunofluorescence assay further demonstrated that G1 VP7 polyclonal antibody can bind to different RVAs, including Wa (genotype G1P[8]), DS-1(genotype G2P[4]), SA11 (genotype G3P[2]), and human G9P[8] RV strains. In addition, double sandwich ELISA showed that VP7 polyclonal antibody could be used to detect rotavirus in clinical samples.Conclusions:The soluble G1 VP7 protein was successfully expressed and VP7 antibody was obtained. The G1 VP7 polyclonal antibody could bind to a variety of G-type rotaviruses, which lays a foundation for the establishment of detection method of different G type rotaviruses.
