Purification of foot-and-mouth disease virus by heparin as ligand for certain strains

被引:16
作者
Du, Ping [1 ]
Sun, Shiqi [1 ]
Dong, Jinjie [1 ]
Zhi, Xiaoying [1 ]
Chang, Yanyan [1 ]
Teng, Zhidong [1 ]
Guo, Huichen [1 ]
Liu, Zaixin [1 ]
机构
[1] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Natl Foot & Mouth Dis Reference Lab, State Key Lab Vet Etiol Biol, Xujiaping 1, Lanzhou 730046, Gansu, Peoples R China
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2017年 / 1049卷
关键词
Foot-and-mouth disease virus; Heparin affinity chromatography; Anion exchange; Virus purification; B SURFACE-ANTIGEN; AFFINITY-CHROMATOGRAPHY; EMPTY PARTICLES; INFECTION; RECEPTOR; SYSTEM; CELLS;
D O I
10.1016/j.jchromb.2016.12.033
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The goal of this project was to develop an easily operable and scalable process for the recovery and purification of foot-and-mouth disease virus (FMDV) from cell culture. Heparin resins HipTrap Heparin HP and AF-Heparin HC-650 were utilized to purify FMDV O/HN/CHA/93. Results showed that the purity of AF-Heparin HC-650 was ideal. Then, the O/HN/CHA/93, O/Tibet/CHA/99, Asia I/FIN/06, and A/CHA/HB/2009 strains were purified by AF-Heparin HC-650. Their affinity/virus recoveries were approximately 51.2%/45.8%, 71.5%/70.9%, 96.4%/73.5, and 59.5%/42.1%, respectively. During a stepwise elution strategy, the viral particles were mainly eluted at 300 mM ionic strength peaks. The heparin affinity chromatography process removed more than 94% of cellular and medium proteins. Anion exchange resin Capto Q captured four FMD virus particles; 40% of binding proteins and 80%-90% of viral particles were eluted at 450 mM NaCI. Moreover, ionic strength varied from 30 to 450 mM had no effect on the immunity to FMDV. The results revealed that heparin sulfate may be the main receptor for CHA/99 strain attachment-susceptible cells. Heparin affinity chromatography can reach perfect results, especially when used as a ligand of the virus. Anion exchange is useful only as previous step for further purification. 2016 Published by Elsevier B.V.
引用
收藏
页码:16 / 23
页数:8
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