Poliovirus separation from cell extracts using capillary electrophoresis: potential use in vaccine production and control?
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.
Department of Analytical Chemistry and Pharmaceutical Technology, Vrije Universiteit Brussel-VUB, Center for Pharmaceutical Research-CePhaR, Laarbeeklaan 103, B-1090, Brussels, Belgium.
Iuliana
I
Oita
Hadewych
H
Halewyck
Sigrid
S
Pieters
Bert
B
Thys
Yvan Vander
YV
Heyden
Bart
B
Rombaut
D057205:Automation, Laboratory / Q000379:methods; D002457:Cell Extracts / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D006801:Humans; D017955:Poliovirus / Q000302:isolation & purification*; D023321:Poliovirus Vaccines / Q000302:isolation & purification*; D013678:Technology, Pharmaceutical / Q000379:methods*; D013997:Time Factors; D019562:Viral Load / Q000379:methods*
D002457:Cell Extracts; D023321:Poliovirus Vaccines