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Isolating Pathogen-Specific Human Monoclonal Antibodies (hmAbs) Using Bacterial Whole Cells as Molecular Probes

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Vaccine Delivery Technology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2183))

Abstract

The immunoglobulin capture assay (ICA) enables the enrichment for pathogen-specific plasmablasts from individuals with a confirmed adaptive immune response to vaccination or disseminated infection. Only single recombinant antigens have been used previously as probes in this ICA and it was unclear whether the method was applicable to complex probes such as whole bacterial cells. Here, we describe the enrichment of plasmablasts specific for polysaccharide and protein antigens of both Streptococcus pneumoniae and Neisseria meningitidis using whole formalin-fixed bacterial cells as probes. The modified ICA protocol described here allowed for a pathogen-specific hmAb cloning efficiency of >80%.

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Acknowledgments

This study was funded, in part, by research grants from John and Michelle Bresnahan via MeningitisNow (awarded to P.R.L), an Imperial College Confidence-in-Concept grant (Number PS3075_WMNP awarded to P.R.L and F.A.B), a UK Biotechnology and Biological Sciences Research Council PhD studentship (BB/R505742/1 awarded to C.A.G), and a UK Medical Research Council Career Development Award (MR/S007490/1 awarded to F.A.B).

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Author notes

  1. Sara Siris and Camilla Gladstone contributed equally to this work.Paul Langford and Fadil Bidmos contributed equally to this work.

Authors and Affiliations

  1. Section of Paediatric Infectious Disease, Department of Infectious Disease, Imperial College London, London, UK

    Sara Siris, Camilla A. Gladstone, Paul R. Langford & Fadil A. Bidmos

  2. Flow Cytometry Core Facility, National Heart and Lung Institute, Imperial College London, London, UK

    Yanping Guo

  3. Section of Immunology of Infection, Department of Infectious Disease, Imperial College London, London, UK

    Christopher L. Pinder, Robin J. Shattock & Paul F. McKay

Authors
  1. Sara Siris
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  2. Camilla A. Gladstone
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  3. Yanping Guo
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  4. Christopher L. Pinder
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  5. Robin J. Shattock
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  6. Paul F. McKay
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  7. Paul R. Langford
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  8. Fadil A. Bidmos
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Corresponding author

Correspondence to Fadil A. Bidmos .

Editor information

Editors and Affiliations

  1. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, NY, USA

    Blaine A. Pfeifer

  2. Department of Chemical and Biological Engineering, University at Buffalo, The State University of New York, Buffalo, NY, USA

    Andrew Hill

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Siris, S. et al. (2021). Isolating Pathogen-Specific Human Monoclonal Antibodies (hmAbs) Using Bacterial Whole Cells as Molecular Probes. In: Pfeifer, B.A., Hill, A. (eds) Vaccine Delivery Technology. Methods in Molecular Biology, vol 2183. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0795-4_2

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  • DOI: https://doi.org/10.1007/978-1-0716-0795-4_2

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0794-7

  • Online ISBN: 978-1-0716-0795-4

  • eBook Packages: Springer Protocols

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