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Anal Chem. 2013 Jun 18;85(12):6089-94. doi: 10.1021/ac400968q. Epub 2013 May 28.

Isoelectric point determination of live polioviruses by capillary isoelectric focusing with whole column imaging detection.

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  • 1Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands. yvonne.thomassen@intravacc.nl

Abstract

Using a capillary isoelectric focusing-whole column imaging detection (CIEF-WCID) method, the isoelectric points (pI) of complete intact polioviruses were determined. The polioviruses that were analyzed are the commonly used viruses for the production of inactivated polio vaccines (IPV)-Mahoney (type 1), MEF (type 2), and Saukett (type 3)-as well as for attenuated oral polio vaccines (OPV) and Sabin types 1, 2, and 3. A method for analyzing biological hazardous components (biological safety level 2) was set up for the CIEF-WCID analyzer used. This method is based on closed circuits. The determined pI's were 6.2 for Mahoney, 6.7 for MEF-1, and 5.8 for Saukett. The pI's of Sabin types 1, 2, and 3 viruses were 7.4, 7.2, and 6.3, respectively. Resolution of the virus peaks was shown to be reproducible. Using this adjusted CIEF-WCID technique, the pI of biologically hazardous components like toxins or viruses can be determined, which is beneficial for the development of vaccine production methods among others.

PMID:
23672432
[PubMed - indexed for MEDLINE]
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