Smales, C.M. and Pepper, D.S. and James, D.C. (2001) Evaluation of Protein Modification during Anti-Viral Heat Bioprocessing by Electrospray Ionization Mass Spectrometry. Rapid Communications in Mass Spectrometry, 15 (5). pp. 351-356. ISSN 0951-4198. (The full text of this publication is not available from this repository)
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During the preparation of therapeutic plasma and recombinant protein biopharmaceuticals heat-treatment is routinely applied as a means of viral inactivation. However, as most proteins denature and aggregate under heat stress, it is necessary to add thermostabilizing excipients to protein formulations destined for anti-viral heat-treatment in order to prevent protein damage. Anti-viral heat-treatment bioprocessing therefore requires that a balance be found between the bioprocessing conditions, virus kill and protein integrity. In this study we have utilized a simple model protein, beta -lactoglobulin, to investigate the relationship between virucidal heat-treatment conditions (protein formulation and temperature) and the type and extent of protein modification in the liquid state. A variety of industrially relevant heat-treatments were undertaken, using formulations that included sucrose as a thermostabilizing excipient. Using liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) we show here that protein mo difications do occur with increasingly harsh heat-treatment. The predominant modification under these conditions was protein glycation by either glucose or fructose derived from hydrolyzed sucrose. Advanced glycation end products and additional unidentified products were also present in beta -lactoglobulin protein samples subjected to extended heat-treatment. These findings have implications for the improvement of anti-viral heat-treatment bioprocesses to ensure the safety and efficacy of protein biopharmaceuticals. Copyright (C) 2001 John Wiley & Sons, Ltd.
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||Mark Smales|
|Date Deposited:||28 Oct 2008 19:28|
|Last Modified:||14 Jan 2010 14:23|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/6210 (The current URI for this page, for reference purposes)|