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Impact of the sample matrix composition on the signal enhancement in the capillary electrophoretic separation of poliovirus samples
Authors:Iuliana Oita  Hadewych Halewyck  Bert Thys  Bart Rombaut  Yvan Vander Heyden
Institution:1. Vrije Universiteit Brussel-VUB, Center for Pharmaceutical Research-CePhaR, Department of Analytical Chemistry and Pharmaceutical Technology, Laarbeeklaan 103, B-1090, Brussels, Belgium;2. Vrije Universiteit Brussel-VUB, Center for Pharmaceutical Research-CePhaR, Department of Pharmaceutical Biotechnology & Molecular Biology, Laarbeeklaan 103, B-1090, Brussels, Belgium
Abstract:The development of capillary electrophoretic applications aiming to provide reliable stability assessment of viral suspensions, to detect subviral particles from cell extracts or to study the interactions between virus particles and various biomolecules, cannot be done without a thorough understanding of the sample matrix contribution to the observed electrophoretic behaviour. The present study thoroughly investigates the effect of the sample matrix on the electrophoretic behaviour of poliovirus injected as sample plugs of 1%, 5% and 12% effective capillary length. The effect of the sample matrix for three different poliovirus batches was evaluated. Additionally, simulated samples, obtained from concentrated poliovirus suspensions of high purity and diluted with commonly used lab buffers in order to obtain samples with either high or low conductivities, were also investigated. The goal of the study was to obtain a better understanding of the effect of the sample matrix on the signal enhancement, in order to define a general approach allowing a repeatable capillary electrophoretic (CE) separation of poliovirus from complex samples. This study clearly demonstrates that the sample matrix has an important influence on the sensitivity of the CE poliovirus separations. Translation of these observations into routine practice involves several compromises and a set of rules in order to reduce day-to-day variation and to maximize sensitivity.
Keywords:Capillary electrophoresis  Poliovirus  Sensitivity improvement  Signal enhancement  Virus quantitation
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