Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands.
Anal Chem. 2013 Jun 18;85(12):6089-94. doi: 10.1021/ac400968q. Epub 2013 May 28.
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.
采用毛细管等电聚焦-全长柱成像检测(CIEF-WCID)方法,测定了完整的脊髓灰质炎病毒的等电点(pI)。分析的脊髓灰质炎病毒包括常用于生产灭活脊髓灰质炎疫苗(IPV)的病毒——马霍尼(类型 1)、MEF(类型 2)和萨基特(类型 3),以及减毒口服脊髓灰质炎疫苗(OPV)和萨宾 1、2 和 3 型。为使用的 CIEF-WCID 分析仪建立了一种用于分析生物危害成分(生物安全 2 级)的方法。该方法基于闭路系统。测定的 pI 值分别为马霍尼 6.2、MEF-1 6.7 和萨基特 5.8。萨宾 1、2 和 3 型病毒的 pI 值分别为 7.4、7.2 和 6.3。病毒峰的分辨率显示可重复。使用这种调整后的 CIEF-WCID 技术,可以测定毒素或病毒等生物危害成分的 pI 值,这有利于疫苗生产方法等方面的发展。