New capillary gel electrophoresis method for fast and accurate identification and quantification of multiple viral proteins in influenza vaccines.
作者： Ewoud van Tricht , Lars Geurink , Bojana Pajic , Johan Nijenhuis , Harold Backus
DOI: 10.1016/J.TALANTA.2015.07.047
关键词: Capillary electrophoresis 、 Accuracy and precision 、 Influenza A virus 、 Hemagglutinin (influenza) 、 Virosome 、 Protein purification 、 Chemistry 、 Chromatography 、 Analyte 、 Sample preparation
摘要: Current methods for the identification and/or quantification of viral proteins in influenza virus and virosome samples suffer from long analysis times, limited protein coverage low accuracy precision. We studied optimized capillary gel electrophoresis (CGE) order to achieve faster enhanced characterization proteins. Sample preparation as well composition buffer was investigated adequate separation relatively short times. The total sample (reduction deglycosylation) could be carried out efficiently within two hours. Hydrodynamic injection, voltage, temperature were full factorial design. final method validated showed good performance hemagglutinin fragment 1 (HA1), 2 (HA2), matrix (M) nucleoprotein (NP). CGE allowed different strains based on their specific profile. B/Brisbane inactivated analyzed one day. results (titers) comparable single radial immune-diffusion (SRID), but has advantage a much time results. A/Christchurch upstream process demonstrated applicability high complexity. used same analyte concentration range RP-HPLC method, better precision accuracy. Overall, shorter, allowing 100 4 days instead 10 SRID.
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