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Evaluation of Protein Modification during Anti-Viral Heat Bioprocessing by Electrospray Ionization Mass Spectrometry

Smales, Christopher Mark, Pepper, Duncan S., James, David 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. (doi:10.1002/rcm.232) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:6210)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided.
Official URL:
http://dx.doi.org/10.1002/rcm.232

Abstract

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.

Item Type: Article
DOI/Identification number: 10.1002/rcm.232
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Mark Smales
Date Deposited: 28 Oct 2008 19:28 UTC
Last Modified: 05 Nov 2024 09:38 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/6210 (The current URI for this page, for reference purposes)

University of Kent Author Information

Smales, Christopher Mark.

Creator's ORCID: https://orcid.org/0000-0002-2762-4724
CReDIT Contributor Roles:

James, David C..

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