Toolbox for non-intrusive structural and functional analysis of recombinant VLP based vaccines

A case study with hepatitis B vaccine

Anke Mulder, Bridget Carragher, Victoria Towne, Yuan Meng, Yang Wang, Lance Dieter, Clinton S. Potter, Michael W. Washabaugh, Robert D. Sitrin, Qinjian Zhao

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Background: Fundamental to vaccine development, manufacturing consistency, and product stability is an understanding of the vaccine structure-activity relationship. With the virus-like particle (VLP) approach for recombinant vaccines gaining popularity, there is growing demand for tools that define their key characteristics. We assessed a suite of non-intrusive VLP epitope structure and function characterization tools by application to the Hepatitis B surface antigen (rHBsAg) VLP-based vaccine. Methodology: The epitope-specific immune reactivity of rHBsAg epitopes to a given monoclonal antibody was monitored by surface plasmon resonance (SPR) and quantitatively analyzed on rHBsAg VLPs in-solution or bound to adjuvant with a competitive enzyme-linked immunosorbent assay (ELISA). The structure of recombinant rHBsAg particles was examined by cryo transmission electron microscopy (cryoTEM) and in-solution atomic force microscopy (AFM). Principal Findings: SPR and competitive ELISA determined relative antigenicity in solution, in real time, with rapid turn-around, and without the need of dissolving the particulate aluminum based adjuvant. These methods demonstrated the nature of the clinically relevant epitopes of HBsAg as being responsive to heat and/or redox treatment. In-solution AFM and cryoTEM determined vaccine particle size distribution, shape, and morphology. Redox-treated rHBsAg enabled 3D reconstruction from CryoTEM images - confirming the previously proposed octahedral structure and the established lipid-to-protein ratio of HBsAg particles. Results from these non-intrusive biophysical and immunochemical analyses coalesced into a comprehensive understanding of rHBsAg vaccine epitope structure and function that was important for assuring the desired epitope formation, determinants for vaccine potency, and particle stability during vaccine design, development, and manufacturing. Significance: Together, the methods presented here comprise a novel suite of non-intrusive VLP structural and functional characterization tools for recombinant vaccines. Key VLP structural features were defined and epitope-specific antigenicity was quantified while preserving epitope integrity and particle morphology. These tools should facilitate the development of other VLP-based vaccines.

Original languageEnglish (US)
Article numbere33235
JournalPLoS One
Volume7
Issue number4
DOIs
StatePublished - Apr 6 2012
Externally publishedYes

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Virus-Like Particle Vaccines
Hepatitis B Vaccines
Functional analysis
Synthetic Vaccines
virus-like particles
hepatitis B
Viruses
Structural analysis
epitopes
Epitopes
Vaccines
vaccines
case studies
Virion
Hepatitis B Surface Antigens
Vaccine Potency
Cryoelectron Microscopy
surface plasmon resonance
Immunosorbents
atomic force microscopy

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Toolbox for non-intrusive structural and functional analysis of recombinant VLP based vaccines : A case study with hepatitis B vaccine. / Mulder, Anke; Carragher, Bridget; Towne, Victoria; Meng, Yuan; Wang, Yang; Dieter, Lance; Potter, Clinton S.; Washabaugh, Michael W.; Sitrin, Robert D.; Zhao, Qinjian.

In: PLoS One, Vol. 7, No. 4, e33235, 06.04.2012.

Research output: Contribution to journalArticle

Mulder, A, Carragher, B, Towne, V, Meng, Y, Wang, Y, Dieter, L, Potter, CS, Washabaugh, MW, Sitrin, RD & Zhao, Q 2012, 'Toolbox for non-intrusive structural and functional analysis of recombinant VLP based vaccines: A case study with hepatitis B vaccine', PLoS One, vol. 7, no. 4, e33235. https://doi.org/10.1371/journal.pone.0033235
Mulder, Anke ; Carragher, Bridget ; Towne, Victoria ; Meng, Yuan ; Wang, Yang ; Dieter, Lance ; Potter, Clinton S. ; Washabaugh, Michael W. ; Sitrin, Robert D. ; Zhao, Qinjian. / Toolbox for non-intrusive structural and functional analysis of recombinant VLP based vaccines : A case study with hepatitis B vaccine. In: PLoS One. 2012 ; Vol. 7, No. 4.
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