A Combined LC-MS and Immunoassay Approach to Characterize Preservative-Induced Destabilization of Human Papillomavirus Virus-like Particles Adsorbed to an Aluminum-Salt Adjuvant
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Issue Date
2024-07-02Author
Caringal, Ria T.
Hickey, John M.
Sharma, Nitya
Jerajani, Kaushal
Bewaji, Oluwadara
Brendle, Sarah
Christensen, Neil
Batwal, Saurabh
Mahedvi, Mustafa
Rao, Harish
Dogar, Vikas
Chandrasekharan, Rahul
Shaligram, Umesh
Joshi, Sangeeta B.
Volkin, David B.
Publisher
MDPI
Type
Article
Article Version
Scholarly/refereed, publisher version
Rights
Copyright © 2024 by the authors.
Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
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During the multi-dose formulation development of recombinant vaccine candidates, protein antigens can be destabilized by antimicrobial preservatives (APs). The degradation mechanisms are often poorly understood since available analytical tools are limited due to low protein concentrations and the presence of adjuvants. In this work, we evaluate different analytical approaches to monitor the structural integrity of HPV16 VLPs adsorbed to Alhydrogel™ (AH) in the presence and absence of APs (i.e., destabilizing m-cresol, MC, or non-destabilizing chlorobutanol, CB) under accelerated conditions (pH 7.4, 50 °C). First, in vitro potency losses displayed only modest correlations with the results from two commonly used methods of protein analysis (SDS-PAGE, DSC). Next, results from two alternative analytical approaches provided a better understanding of physicochemical events occurring under these same conditions: (1) competitive ELISA immunoassays with a panel of mAbs against conformational and linear epitopes on HPV16 VLPs and (2) LC-MS peptide mapping to evaluate the accessibility/redox state of the 12 cysteine residues within each L1 protein comprising the HPV16 VLP (i.e., with 360 L1 proteins per VLP, there are 4320 Cys residues per VLP). These methods expand the limited analytical toolset currently available to characterize AH-adsorbed antigens and provide additional insights into the molecular mechanism(s) of AP-induced destabilization of vaccine antigens.
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Caringal RT, Hickey JM, Sharma N, Jerajani K, Bewaji O, Brendle S, Christensen N, Batwal S, Mahedvi M, Rao H, Dogar V, Chandrasekharan R, Shaligram U, Joshi SB, Volkin DB. A Combined LC-MS and Immunoassay Approach to Characterize Preservative-Induced Destabilization of Human Papillomavirus Virus-like Particles Adsorbed to an Aluminum-Salt Adjuvant. Vaccines (Basel). 2024 May 26;12(6):580. doi: 10.3390/vaccines12060580. PMID: 38932309; PMCID: PMC11209183.
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Except where otherwise noted, this item's license is described as: Copyright © 2024 by the authors.
Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).