Poliovirus separation from cell extracts using capillary electrophoresis: Potential use in vaccine production and control?

被引:13
作者
Oita, Iuliana [2 ]
Halewyck, Hadewych [1 ]
Pieters, Sigrid [2 ]
Thys, Bert [1 ]
Heyden, Yvan Vander [2 ]
Rombaut, Bart [1 ]
机构
[1] VUB, Dept Pharmaceut Biotechnol & Mol Biol, Ctr Pharmaceut Res CePhaR, B-1090 Brussels, Belgium
[2] VUB, Ctr Pharmaceut Res CePhaR, Dept Analyt Chem & Pharmaceut Technol, B-1090 Brussels, Belgium
关键词
Capillary electrophoresis; Poliovirus; Sensitivity improvement; Signal enhancement; Virus detection; Virus quantitation; VIRUS; SENSITIVITY; ELECTROMIGRATION; ASSAY; ELECTROCHROMATOGRAPHY; PURIFICATION; ABSORBENCY; PRINCIPLES; MICROCHIPS; PARTICLES;
D O I
10.1016/j.jviromet.2012.05.015
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Rapid assessment of the concentration of virus particles in a given sample remains a challenge. Modern separation methods, such as capillary electrophoresis, were proposed recently to study viruses and viral infection or to separate and characterize viral vaccines in a time-efficient manner. Even though capillary electrophoresis is much more rapid than traditional virological methods and has the advantages of automation, increased precision and reliability, it has the drawback of reduced sensitivity for low concentrations. A sensitivity improvement is then necessary in many cases for a successful application. However, to date, only highly purified viral samples were examined using capillary electrophoresis. The injection of larger sample volumes, followed by intra-capillary concentration, was used in this study for cell extracts. Poliovirus was successfully detected rapidly, without any laborious staining procedures and incubation times. The method is simple, fast, automatic, requires only minute amounts of samples and reagents, and no expensive dyes or biological reagents. Additionally, the method showed a potential for monitoring the viral load during growth and purification, with obvious prospects for the optimization of the variable and time-consuming virus propagation procedures. The results of this study provide a potential basis for the development of routine methods for viral particles analysis, irrespective of their infective properties. In the future, the capillary electrophoresis test could help study the relationship between the intact poliovirus particles and the D-antigenic properties of a viral suspension, or could represent a supplementary or alternative test for virus concentration and D-antigen assays during vaccine production. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:7 / 17
页数:11
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