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

文献类型： 外文期刊

第一作者： Du, Ping

作者： Du, Ping;Sun, Shiqi;Dong, Jinjie;Zhi, Xiaoying;Chang, Yanyan;Teng, Zhidong;Guo, Huichen;Liu, Zaixin

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关键词： Foot-and-mouth disease virus;Heparin affinity chromatography;Anion exchange;Virus purification

期刊名称：JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES （ 影响因子：3.205； 五年影响因子：3.068 ）

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收录情况： SCI

摘要： Highlights ? FMD strains displayed different affinities to heparin resins, heparin affinity chromatography would reach perfect results only when heparin was ligand of virus，and CHA/99 strain may be utilized heparin sulfate as the main receptor to attach susceptible cells. ? The purified results of anion exchange were not affected by FMD strain divergence. ? Ionic strength varied from 30 to 450 mM had no effect on immunity of FMDV. Abstract 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/HN/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 300mM 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 450mM NaCl. Moreover, ionic strength varied from 30 to 450mM 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. ]]>

分类号： Q503

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