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Use of Chlamydial Elementary Bodies as Probes to Isolate Pathogen-Specific Human Monoclonal Antibodies

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

Abstract

Chlamydia trachomatis is one of the most prevalent sexually transmitted infectious agents in the world and the leading cause of infectious blindness. The role of antibodies in the prevention and clearance of infection is still not fully understood, but the analysis of the immunoglobulin response to novel vaccine candidates is an important part of many of these studies. In this chapter, we describe a novel method to identify and isolate Chlamydia-specific memory B cells by fluorescence-activated cell sorting (FACS) using fluorescently labeled whole bacteria from cryopreserved human PBMC samples. This method allows for live single cells to be sorted for cell culture, in vitro assays, single-cell RNA sequencing, and cloning of paired heavy and light chains for recombinant monoclonal antibody production.

Key words

  • Antigen-specific
  • B cell
  • Antibody
  • Chlamydia
  • Elementary body
  • Immunoglobulin
  • IgG

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Acknowledgments

We would like to thank Prof. Martin Holland and Dr. Harry Pickering of the London School of Hygiene and Tropical Medicine, and Drs. Frank Follmann, Ida Rosenkrands, and Anja Olsen of the Statens Serum Institut for providing the purified elementary bodies used for this work.

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Correspondence to Paul F. McKay .

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Pinder, C.L., McKay, P.F., Shattock, R.J. (2021). Use of Chlamydial Elementary Bodies as Probes to Isolate Pathogen-Specific Human Monoclonal Antibodies. 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_3

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  • DOI: https://doi.org/10.1007/978-1-0716-0795-4_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0794-7

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