Purification of H5N1 influenza virus by different chromatography media
10.3760/cma.j.cn112309-20221010-00326
- VernacularTitle:不同层析介质纯化H5N1型流感病毒的研究
- Author:
Bo LIU
1
;
Jiayou ZHANG
;
Fang LI
;
Zhegang ZHANG
;
Yaqi JI
;
Rong ZHOU
;
Xuedan LI
;
Ze LI
;
Qingda LI
;
Wenyi WU
;
Xiaoming YANG
Author Information
1. 武汉生物制品研究所有限责任公司病毒性疫苗研究二室，武汉 430207
- Keywords:
H5N1 influenza virus;
Gel filtration chromatography medium;
Capto Core700;
Mixed-mode chromatography medium
- From:
Chinese Journal of Microbiology and Immunology
2023;43(3):217-221
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To purify H5N1 influenza virus concentrate prepared by MDCK cells with a new mixed-mode chromatography medium Capto Core700 and the traditional medium Sepharose 4FF, and to compare the separation and purification efficacy of the two media.Methods:Capto Core700 and Sepharose 4FF were used to purify inactivated H5N1 influenza virus concentrate. The morphology of virus particles in different samples was then observed under a transmission electron microscope. Single radial immunodiffusion (SRID), Folin-Phenol (Lowry) method, double-antibody sandwich ELISA and qPCR were used to detect hemagglutinin, total protein, host cell protein (HCP) and host cell DNA (HCD) before and after purification. The recovery rate of virus antigen and the removal rate of impurities were calculated. The immunogenicity of the viruses purified with different media was analyzed using animal experiments. Difference in the purification efficacy of the two chromatography media was analyzed by t-test. Results:H5N1 influenza viruses purified by Capto Core700 or Sepharose 4FF showed the typical influenza virus morphology under transmission electron microscope. There was no significant difference in the recovery rate of hemagglutinin between the two chromatography media ( P>0.05), but compared with Sepharose 4FF, Capto Core700 had a higher removal rate of impurities (total protein, HCP, HCD) and the difference was statistically significant ( P<0.05). Animal experiments showed that the viruses purified by the two chromatography media had good immunogenicity. Conclusions:Compared with Sepharose 4FF chromatography medium, Capto Core700 could more effectively remove process-related impurities such as HCP, HCD and total protein without affecting the recovery rate of viral antigen. This study provided reference for the development of purification technology in the production of H5N1 influenza virus vaccine in MDCK cells.
