Molecular and structural characterization of the L1 virus-like particles that are used as vaccine antigens in<i>Cervarix</i>™, the AS04-adjuvanted HPV-16 and -18 cervical cancer vaccine

Deschuyteneer, M; Elouahabi, Abdelatif; Plainchamp, Dominique; Plisnier, Michel; Soete, Dominique; Corazza, Yvon; Lockman, Laurence; Giannini, Sandra L.; Deschamps, Marguerite

doi:10.4161/hv.6.5.11023

Cited by 122 publications

(92 citation statements)

References 41 publications

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“…Monoclonal antibodies H16.V5, H16.1A, H16.14J, and H263.A2 examined in the previous study all target conformational epitopes located on combinations of the apical surface-exposed loops of L1 proteins. However, a novel neutralizing monoclonal antibody, H16.U4, generated against HPV16 L1 VLP in an earlier study (22) bound capsids differently (21,23), albeit with a weaker neutralizing ability (13,21,(23)(24)(25) than those of the four previously studied MAbs. A mutational analysis mapped the binding site of H16.U4 to amino acids 427 to 445 of the C-terminal arm of L1 in the canyon between capsomers (21).…”

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confidence: 99%

“…Since the HPV life cycle depends on the differentiation of keratinocytes, it is difficult to purify high-titer virus stocks for structural studies. Virus-like particles (VLPs) that are devoid of viral genome (11) have been used successfully for structural studies (8,12,13), whereas both pseudovirus (PsV) and quasivirus (QV), which contain expression plasmid DNA (14,15), have been used for structural studies and infectivity assays (9,10). For the work presented here, quasivirus has been used throughout.…”

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confidence: 99%

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The U4 Antibody Epitope on Human Papillomavirus 16 Identified by Cryo-electron Microscopy

Guan

Bywaters

Brendle

et al. 2015

J Virol

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The human papillomavirus (HPV) major structural protein L1 composes capsomers that are linked together through interactions mediated by the L1 C terminus to constitute a T‫7؍‬ icosahedral capsid. H16.U4 is a type-specific monoclonal antibody recognizing a conformation-dependent neutralizing epitope of HPV thought to include the L1 protein C terminus. The structure of human papillomavirus 16 (HPV16) complexed with H16.U4 fragments of antibody (Fab) was solved by cryo-electron microscopy (cryo-EM) image reconstruction. Atomic structures of virus and Fab were fitted into the corresponding cryo-EM densities to identify the antigenic epitope. The antibody footprint mapped predominately to the L1 C-terminal arm with an additional contact point on the side of the capsomer. This footprint describes an epitope that is presented capsid-wide. However, although the H16.U4 epitope suggests the presence of 360 potential binding sites exposed in the capsid valley between each capsomer, H16.U4 Fab bound only to epitopes located around the icosahedral five-fold vertex of the capsid. Thus, the binding characteristics of H16.U4 defined in this study showed a distinctive selectivity for local conformation-dependent interactions with specific L1 invading arms between five-fold related capsomers. H uman papillomavirus (HPV) infections continue to be a significant health burden in patient populations (1, 2). Although commercial vaccines targeting the viral capsid proteins have been applied successfully to protect against high-risk HPV, the efficacy of vaccines is genotype specific, and vaccines provide little therapeutic benefit against existing infections (3). Understanding the antigenic nature of the HPV capsid offers an opportunity to discover structural features that are crucial to capsid integrity and conserved across species. Panels of monoclonal antibodies and mutational analyses have helped to define several antigenic epitopes (4-10); however, determining the conformational epitopes on the capsid surface requires structural analyses, which can be accomplished by cryo-electron microscopy (cryo-EM) technology. Since the HPV life cycle depends on the differentiation of keratinocytes, it is difficult to purify high-titer virus stocks for structural studies. Virus-like particles (VLPs) that are devoid of viral genome (11) have been used successfully for structural studies (8, 12, 13), whereas both pseudovirus (PsV) and quasivirus (QV), which contain expression plasmid DNA (14, 15), have been used for structural studies and infectivity assays (9, 10). For the work presented here, quasivirus has been used throughout.Papillomaviruses form a nonenveloped Tϭ7 icosahedral capsid that is ϳ55 to 60 nm in diameter and contains a circular double-stranded DNA (dsDNA) genome of 8 kb. The capsid is comprised of 360 copies of the L1 major structural protein and an uncertain number of the L2 minor structural protein (15,16). Five copies of the L1 protein intertwine to form each capsomer, and 72 capsomers interact to constitute a capsid. T...

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confidence: 99%

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confidence: 99%

The U4 Antibody Epitope on Human Papillomavirus 16 Identified by Cryo-electron Microscopy

Guan

Bywaters

Brendle

et al. 2015

J Virol

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“…Several VLP products have been evaluated in clinical trials. In addition to a hepatitis B virus VLP-based licensed vaccine, two human papillomavirus (HPV) VLP vaccines were recently licensed in the United States (6,9,15,38).…”

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confidence: 99%

H5N1 Virus-Like Particle Vaccine Elicits Cross-Reactive Neutralizing Antibodies That Preferentially Bind to the Oligomeric Form of Influenza Virus Hemagglutinin in Humans

Khurana

Verma

et al. 2011

J Virol

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“…This is also a common practice in analytics for human vaccines. [30][31][32][33][34][35] Among the mAbs used for the HEV antigen, the most critical mAb is a protective and neutralizing antibody, mAb 8C11. MAb 8C11 recognizes conformational epitopes that are composed of 3 discontinuous peptide segments in a dimeric form of the truncated HEV capsid protein.…”

Section: Discussionmentioning

confidence: 99%

“…Using multiple epitope approach increases the confidence in antigen characterization and in assuring product consistency as in the case for HPV vaccines. [32][33][34] The higher detection limit of the blocking rate was identified as 95% because the lower limit of ELISA value is 0.05, and the value of no blocking group was controlled between 1.0 and 1.5. Three more level of blocking rates (50%, 75% and 90%) identified the relationship between each blocking and detection antibody (Fig.…”

Section: Immunochemical Methodsmentioning

confidence: 99%

Comparable quality attributes of hepatitis E vaccine antigen with and without adjuvant adsorption-dissolution treatment

Zhang

Yang

et al. 2015

Human Vaccines & Immunotherapeutics

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Most vaccines require adjuvants for antigen stabilization and immune potentiation. Aluminum-based adjuvants are the most widely used adjuvants for human vaccines. Previous reports demonstrated the preservation of antigen conformation and other antigen characteristics after recovery from adjuvanted Hepatitis B and human papillomavirus vaccines. In this study, we used a combination of various physiochemical and immunochemical methods to analyze hepatitis E vaccine antigen quality attributes after recovery from adjuvants. All biochemical and biophysical methods showed similar characteristics of the p239 protein after recovery from adjuvanted vaccine formulation compared to the antigen in solution which never experienced adsorption/desorption process. Most importantly, we demonstrated full preservation of key antigen epitopes post-recovery from adjuvanted vaccine using a panel of murine monoclonal antibodies as exquisite probes. Antigenicity of p239 was probed with a panel of 9 mAbs using competition/blocking ELISA, surface plasmon resonance and sandwich ELISA methods. These multifaceted analyses demonstrated the preservation of antigen key epitopes and comparable protein thermal stability when adsorbed on adjuvants or of the recovered antigen post-dissolution treatment. A better understanding of the antigen conformation in adjuvanted vaccine will enhanced our knowledge of antigen-adjuvant interactions and facilitate an improved process control and development of stable vaccine formulation.

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Molecular and structural characterization of the L1 virus-like particles that are used as vaccine antigens inCervarix™, the AS04-adjuvanted HPV-16 and -18 cervical cancer vaccine

Cited by 122 publications

References 41 publications

The U4 Antibody Epitope on Human Papillomavirus 16 Identified by Cryo-electron Microscopy

The U4 Antibody Epitope on Human Papillomavirus 16 Identified by Cryo-electron Microscopy

H5N1 Virus-Like Particle Vaccine Elicits Cross-Reactive Neutralizing Antibodies That Preferentially Bind to the Oligomeric Form of Influenza Virus Hemagglutinin in Humans

Comparable quality attributes of hepatitis E vaccine antigen with and without adjuvant adsorption-dissolution treatment

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