Document Detail

In-depth process understanding of RECOMBIVAX HB(®) maturation and potential epitope improvements with redox treatment: Multifaceted biochemical and immunochemical characterization.
MedLine Citation:
PMID:  21871939     Owner:  NLM     Status:  Publisher    
Recombinant Hepatitis B surface antigen virus-like particles (VLPs) produced in yeast undergo spontaneous maturation during the vaccine production process, and the biophysical characteristics of the particles with respect to maturation were described in Zhao et al. (2006) [13]. Here we report additional biochemical and immunochemical characterization by various techniques, including the use of a panel of monoclonal antibodies (mAbs) that differ in their selectivity and conformation-sensitivity, for probing surface epitope structures. Crosslinking via interchain disulfide formation and binding of conformational specific antibodies in the mature particles were shown to be progressively enhanced. We show that redox-mediated VLP maturation is superior to heat-induced maturation in terms of generating VLPs which exhibit more complete crosslinking (>95%) and 2- to 3-fold higher antigenicity as defined by conformational antibodies. Therefore, the resulting VLPs from redox treatment resemble more closely their plasma-derived counterparts. The value of using multiple mAbs for probing surface epitopes was clearly demonstrated as different mAbs showed different degrees of sensitivity to the structural changes during HBsAg VLP maturation. The rapid, label-free technology of surface plasmon resonance performed at a single antigen concentration was shown to correlate well with a sandwich ELISA using parallel line analysis, currently implemented for product release and stability testing of RECOMBIVAX HB(®). Surface plasmon resonance offers both convenience and flexibility; multiple mAbs can be tested one at a time in the same set of experiments, providing a means to assess changes to individual epitopes. Taken together, these quantitative analytical tools enable more rapid, in-depth, and comprehensive process monitoring, process optimization, and assessment of product consistency and stability.
Qinjian Zhao; Victoria Towne; Martha Brown; Yang Wang; Dicky Abraham; C Brent Oswald; Juan A Gimenez; Michael W Washabaugh; Ronald Kennedy; Robert D Sitrin
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-8-24
Journal Detail:
Title:  Vaccine     Volume:  -     ISSN:  1873-2518     ISO Abbreviation:  -     Publication Date:  2011 Aug 
Date Detail:
Created Date:  2011-8-29     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8406899     Medline TA:  Vaccine     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2011. Published by Elsevier Ltd.
Merck Research Laboratories, Merck & Co, Inc., West Point, PA 19486, USA.
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