Purification method for cell-cultured influenza virus H5N1
10.3760/cma.j.issn.0254-5101.2019.12.009
- VernacularTitle: 细胞培养流感病毒H5N1的纯化方法研究
- Author:
Zhegang ZHANG
1
;
Dan LUO
2
;
Jiayou ZHANG
2
;
Wei ZHAO
1
;
Ying WANG
1
;
Ran QIU
1
;
Ziyan MENG
1
;
Tian HAN
1
;
Zhiwu XIA
1
;
Changgui LI
3
;
Xiaoming YANG
2
,
4
Author Information
1. The Second Laboratory of Viral Vaccine Research, Wuhan Institute of Biological Products Co., Ltd., Wuhan 430070, China
2. National Engineering Technology Research Center of Combination Vaccines, Wuhan 430060, China
3. Division of Respiratory Virus Vaccines, National Institute for Food and Drug Control, Beijing 100050, China
4. China National Biotechnology Group, Beijing 100029, China
- Publication Type:Journal Article
- Keywords:
Influenza virus;
MDCK cells;
Host cell protein;
Host cell DNA;
Purification
- From:
Chinese Journal of Microbiology and Immunology
2019;39(12):933-936
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To reduce the residual proteins and DNA of host cells in the preparation of H5N1 influenza A virus.
Methods:Core 700 was firstly used to remove residual host cell proteins, and then Capto Q was used to remove host cell DNA. Several batches of H5N1 influenza A virus cultured in Madin-Darby canine kidney (MDCK) cells were purified using this method. The efficiency of purification was evaluated using many methods including quantitative real-time PCR, hemagglutination (HA) test and single radial immunodiffusion assay. Moreover, Benzonase nuclease was used for comparison.
Results:Without the use of Benzonase nuclease, the overall removal rates of host cell DNA and residual proteins were 99.62% and 98.1%, and the HA antigen recovery rate was 66.96%.
Conclusions:This study established a purification strategy with good effect for cell-based influenza vaccines. It can efficiently remove host cell DNA and proteins and achieve a high HA recovery rate. The purification result is no worse than that of adding Benzonase nuclease, suggesting the potential of its application in actual vaccine production.
