Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment

Justin A. Spanier, Daniel R. Frederick, Justin J. Taylor, James R. Heffernan, Dmitri I. Kotov, Tijana Martinov, Kevin C. Osum, Jenna L. Ruggiero, Blake J. Rust, Samuel J. Landry, Marc K. Jenkins, James B. McLachlan and Brian T. Fife ()
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Justin A. Spanier: Center for Immunology, University of Minnesota Medical School
Daniel R. Frederick: Tulane University School of Medicine
Justin J. Taylor: Center for Immunology, University of Minnesota Medical School
James R. Heffernan: Center for Immunology, University of Minnesota Medical School
Dmitri I. Kotov: Center for Immunology, University of Minnesota Medical School
Tijana Martinov: Center for Immunology, University of Minnesota Medical School
Kevin C. Osum: Center for Immunology, University of Minnesota Medical School
Jenna L. Ruggiero: Center for Immunology, University of Minnesota Medical School
Blake J. Rust: Tulane University School of Medicine
Samuel J. Landry: Tulane University School of Medicine
Marc K. Jenkins: Center for Immunology, University of Minnesota Medical School
James B. McLachlan: Tulane University School of Medicine
Brian T. Fife: Center for Immunology, University of Minnesota Medical School

Nature Communications, 2016, vol. 7, issue 1, 1-11

Abstract: Abstract Monoclonal antibodies specific for foreign antigens, auto-antigens, allogeneic antigens and tumour neo-antigens in the context of major histocompatibility complex II (MHCII) are highly desirable as novel immunotherapeutics. However, there is no standard protocol for the efficient generation of monoclonal antibodies that recognize peptide in the context of MHCII, and only a limited number of such reagents exist. In this report, we describe an approach for the generation and screening of monoclonal antibodies specific for peptide bound to MHCII. This approach exploits the use of recombinant peptide:MHC monomers as immunogens, and subsequently relies on multimers to pre-screen and magnetically enrich the responding antigen-specific B cells before fusion and validation, thus saving significant time and reagents. Using this method, we have generated two antibodies enabling us to interrogate antigen presentation and T-cell activation. This methodology sets the standard to generate monoclonal antibodies against the peptide–MHCII complexes.

Date: 2016
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DOI: 10.1038/ncomms11804

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