Assaying the Potency of Influenza Vaccines

Minor, Philip D.

doi:10.3390/vaccines3010090

Cited by 53 publications

(66 citation statements)

References 25 publications

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“…The potential application of cross-reactive nanobodies to pandemic influenza extends beyond immune prophylaxis [ 17 , 64 ], and includes serological surveillance [ 70 ] universal vaccine design [ 71 ] and vaccine potency standardisation [ 72 , 73 ]. The development of cross-reactive monoclonal antibodies to assess structural integrity, quantity and clinical potency of HA in influenza vaccines is of particular interest [ 73 – 75 ]. Influenza viruses have a great capacity to change their genetic material and as such vaccine manufacturers have to update their vaccines every year to accommodate the new circulating strains.…”

Section: Discussionmentioning

confidence: 99%

Cross-Neutralising Nanobodies Bind to a Conserved Pocket in the Hemagglutinin Stem Region Identified Using Yeast Display and Deep Mutational Scanning

Gaiotto

Hufton

2016

PLoS ONE

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Cross-neutralising monoclonal antibodies against influenza hemagglutinin (HA) are of considerable interest as both therapeutics and diagnostic tools. We have recently described five different single domain antibodies (nanobodies) which share this cross-neutralising activity and suggest their small size, high stability, and cleft binding properties may present distinct advantages over equivalent conventional antibodies. We have used yeast display in combination with deep mutational scanning to give residue level resolution of positions in the antibody-HA interface which are crucial for binding. In addition, we have mapped positions within HA predicted to have minimal effect on antibody binding when mutated. Our cross-neutralising nanobodies were shown to bind to a highly conserved pocket in the HA2 domain of A(H1N1)pdm09 influenza virus overlapping with the fusion peptide suggesting their mechanism of action is through the inhibition of viral membrane fusion. We also note that the epitope overlaps with that of CR6261 and F10 which are human monoclonal antibodies in clinical development as immunotherapeutics. Although all five nanobodies mapped to the same highly conserved binding pocket we observed differences in the size of the epitope footprint which has implications in comparing the relative genetic barrier each nanobody presents to a rapidly evolving influenza virus. To further refine our epitope map, we have re-created naturally occurring mutations within this HA stem epitope and tested their effect on binding using yeast display. We have shown that a D46N mutation in the HA2 stem domain uniquely interferes with binding of R2b-E8. Further testing of this substitution in the context of full length purified HA from 1918 H1N1 pandemic (Spanish flu), 2009 H1N1 pandemic (swine flu) and highly pathogenic avian influenza H5N1 demonstrated binding which correlated with D46 whereas binding to seasonal H1N1 strains carrying N46 was absent. In addition, our deep sequence analysis predicted that binding to the emerging H1N1 strain (A/Christchurch/16/2010) carrying the HA2-E47K mutation would not affect binding was confirmed experimentally. This demonstrates yeast display, in combination with deep sequencing, may be able to predict antibody reactivity to emerging influenza strains so assisting in the preparation for future influenza pandemics.

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Section: Discussionmentioning

confidence: 99%

Cross-Neutralising Nanobodies Bind to a Conserved Pocket in the Hemagglutinin Stem Region Identified Using Yeast Display and Deep Mutational Scanning

Gaiotto

Hufton

2016

PLoS ONE

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“…In spite of the advantageous features of the SRID method that have contributed to its continual use as the traditional potency assay for nearly 40 years, there are some significant limitations to the assay that indicate the need for improvement and the development of alternative potency assays 17. In addition to the challenge of timely reagent preparation noted above, other limitations of the SRID include a relatively narrow dynamic range and questionable applicability to some newer types of influenza vaccines being developed.…”

Section: New Assay Developmentmentioning

confidence: 99%

Standardisation of inactivated influenza vaccines—Learning from history

Wood

Weir

2018

Influenza Resp Viruses

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The single radial immunodiffusion assay has been the accepted method for determining the potency of inactivated influenza vaccines since 1978. The worldwide adoption of this assay for vaccine standardisation was facilitated through collaborative studies that demonstrated a high level of reproducibility and its applicability to the different types of influenza vaccine being produced at that time. Clinical evidence indicated the relevance of SRID as a potency assay. Unique features of the SRID assay are likely responsible for its longevity even as newer technologies for vaccine characterisation have been developed and refined. Nevertheless, there are significant limitations to the SRID assay that indicate the need for improvement, and there has been a substantial amount of work undertaken in recent years to develop and evaluate alternative potency assays, including collaborative studies involving research laboratories, regulatory agencies and vaccine manufacturers. Here, we provide an overview of the history of inactivated influenza vaccine potency testing, the current state of alternative assay development and the some of the major challenges to be overcome before implementation of new assays for potency determination.

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“…Specifically, demand is growing for an alternative vaccine potency assay. A well-written and comprehensive review of the current gold-standard method, single radial immunodiffusion (SRID or SRD), was recently published [1]. As mentioned therein, the need for an alternative to SRID is driven by the time delay in flu vaccine development caused by seasonal strain-specific reagents and a growing number of innovative flu vaccine candidates for which SRID is unsuitable.…”

Section: Contents Lists Available At Sciencedirectmentioning

confidence: 99%

Validation of alternative potency assays for influenza vaccines requires clinical studies

Rowlen

2015

Vaccine

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Assaying the Potency of Influenza Vaccines

Cited by 53 publications

References 25 publications

Cross-Neutralising Nanobodies Bind to a Conserved Pocket in the Hemagglutinin Stem Region Identified Using Yeast Display and Deep Mutational Scanning

Cross-Neutralising Nanobodies Bind to a Conserved Pocket in the Hemagglutinin Stem Region Identified Using Yeast Display and Deep Mutational Scanning

Standardisation of inactivated influenza vaccines—Learning from history

Validation of alternative potency assays for influenza vaccines requires clinical studies

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