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采用整柱成像检测的毛细管等电聚焦法测定活脊髓灰质炎病毒的等电点。

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

机构信息

Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands.

出版信息

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

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.

摘要

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。

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