國家衛生研究院 NHRI:Item 3990099045/14481

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Please use this identifier to cite or link to this item: http://ir.nhri.org.tw/handle/3990099045/14481

Title:	Separation and purification of highly infectious enterovirus A71 particles using a strong anion-exchange column
Authors:	Lien, SC;Lu, CC;Shen, YS;Yang, YT;Wu, SR;Fang, CY;Chow, YH;Liao, CL;Chiang, JR;Liu, CC
Contributors:	National Institute of Infectious Diseases and Vaccinology
Abstract:	Virions produced from cell culture is the primary source for production of formalin-inactivated whole virus vaccines for enteroviruses. EV-A71 particles produced from culture system comprise two major types, the immature/empty (E)-particle and the mature/full (F)-particle, which both exhibit low isoelectric point (pI) values but have distinct differences in infectivity and immunogenicity. Although EV-A71 particles can conventionally be separated into E-particle and F-particle using sucrose gradient ultracentrifugation, this procedure is cumbersome and difficult to put into practice for vaccine production. Methods based on ion-exchange chromatography have been exploited to improve the purification efficacy; however, none of them are capable of separating the E- and F-particles efficiently. In this study, we aimed to develop an approach to isolate and purify the highly immunogenic mature EV-A71 particles. By applying a step gradient elution procedure, we successfully isolated the viral structure protein VP0-cleaved particles of EV-A71 from a mixture of cultured viral solution using the Q-membrane anion-exchange chromatography. The elution started with 0.1x phosphate buffered saline (PBS) solution while increasing the percentage of 1x PBS containing 1M NaCl in sequential steps. By this procedure, the VP0-cleaved mature particles and VP0-uncleaved immature particles of EV-A71 could be separated into different fractions in Q-membrane with gradually increased NaCl concentration in elution buffer. The purified VP0-cleaved particles were shown to have characteristics equivalent to those of the highly infectious F-particles of EV-A71. The overall recovery rate for the mature EV-A71 particles by Q-membrane is 56% and its purity was shown to be equivalent to those isolated by the sucrose gradient ultracentrifugation. Our approach provides a simple and efficient purification method for recovering mature, highly infectious virus particles from the EV-A71 culture bulk.
Date:	2022-09-13
Relation:	Journal of Chromatography A. 2022 Sep 13;1680:Article number 463427.
Link to:	http://dx.doi.org/10.1016/j.chroma.2022.463427
JIF/Ranking 2023:	http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=0021-9673&DestApp=IC2JCR
Cited Times(WOS):	https://www.webofscience.com/wos/woscc/full-record/WOS:000874773500008
Cited Times(Scopus):	https://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=85136276182
Appears in Collections:	[Chia-Chyi Liu] Periodical Articles [Ching-Len Liao] Periodical Articles [Joe Yen-Hung Chow] Periodical Articles

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