tetano
Editor, Senior Moderator
Proteomics. 2013 Oct 7. doi: 10.1002/pmic.201300096. [Epub ahead of print]
Surface Modifications of Influenza Proteins Upon Virus Inactivation by Beta-Propiolactone.
She YM, Cheng K, Farnsworth A, Li X, Cyr TD.
Source
Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario, Canada.
Abstract
Inactivation of intact influenza viruses using formaldehyde or beta-propiolactone is essential for vaccine production and safety. The extent of chemical modifications of such reagents on viral proteins needs to be extensively investigated to better control the reactions and quality of vaccines. We have evaluated the effect of beta-propiolactone inactivation on two candidate re-assortant vaccines (NIBRG-121xp and NYMC-X181A) derived from A/California/07/2009 pandemic influenza viruses using high-resolution Fourier transform ion cyclotron mass spectrometry-based proteomic approaches. We report here a UPLC MS/MS method for determining full-length protein sequences of hemagglutinin and neuraminidase through protein delipidation, various enzymatic digestions, and subsequent mass spectrometric analyses of the proteolytic peptides. We also demonstrate the ability to reliably identify hundreds of unique sites modified by propiolactone on the surface of glycoprotein antigens. The location of these modifications correlated in changes to protein folding, conformation and stability, but demonstrated no effect on protein disulfide linkages. In some cases, these modifications resulted in suppression of protein function, an effect that correlated with the degree of change of the modified amino acids' side chain length and polarity. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
Influenza vaccine, Mass spectrometry, Post-translational modification, Virus inactivation, beta-propiolactone
PMID:
24123778
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24123778
Surface Modifications of Influenza Proteins Upon Virus Inactivation by Beta-Propiolactone.
She YM, Cheng K, Farnsworth A, Li X, Cyr TD.
Source
Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, Ontario, Canada.
Abstract
Inactivation of intact influenza viruses using formaldehyde or beta-propiolactone is essential for vaccine production and safety. The extent of chemical modifications of such reagents on viral proteins needs to be extensively investigated to better control the reactions and quality of vaccines. We have evaluated the effect of beta-propiolactone inactivation on two candidate re-assortant vaccines (NIBRG-121xp and NYMC-X181A) derived from A/California/07/2009 pandemic influenza viruses using high-resolution Fourier transform ion cyclotron mass spectrometry-based proteomic approaches. We report here a UPLC MS/MS method for determining full-length protein sequences of hemagglutinin and neuraminidase through protein delipidation, various enzymatic digestions, and subsequent mass spectrometric analyses of the proteolytic peptides. We also demonstrate the ability to reliably identify hundreds of unique sites modified by propiolactone on the surface of glycoprotein antigens. The location of these modifications correlated in changes to protein folding, conformation and stability, but demonstrated no effect on protein disulfide linkages. In some cases, these modifications resulted in suppression of protein function, an effect that correlated with the degree of change of the modified amino acids' side chain length and polarity. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
Influenza vaccine, Mass spectrometry, Post-translational modification, Virus inactivation, beta-propiolactone
PMID:
24123778
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24123778