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

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 monoc...

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Autores principales: Spanier, Justin A., Frederick, Daniel R., Taylor, Justin J., Heffernan, James R., Kotov, Dmitri I., Martinov, Tijana, Osum, Kevin C., Ruggiero, Jenna L., Rust, Blake J., Landry, Samuel J., Jenkins, Marc K., McLachlan, James B., Fife, Brian T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4909947/
https://www.ncbi.nlm.nih.gov/pubmed/27292946
http://dx.doi.org/10.1038/ncomms11804
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author Spanier, Justin A.
Frederick, Daniel R.
Taylor, Justin J.
Heffernan, James R.
Kotov, Dmitri I.
Martinov, Tijana
Osum, Kevin C.
Ruggiero, Jenna L.
Rust, Blake J.
Landry, Samuel J.
Jenkins, Marc K.
McLachlan, James B.
Fife, Brian T.
author_facet Spanier, Justin A.
Frederick, Daniel R.
Taylor, Justin J.
Heffernan, James R.
Kotov, Dmitri I.
Martinov, Tijana
Osum, Kevin C.
Ruggiero, Jenna L.
Rust, Blake J.
Landry, Samuel J.
Jenkins, Marc K.
McLachlan, James B.
Fife, Brian T.
author_sort Spanier, Justin A.
collection PubMed
description 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.
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spelling pubmed-49099472016-06-24 Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment Spanier, Justin A. Frederick, Daniel R. Taylor, Justin J. Heffernan, James R. Kotov, Dmitri I. Martinov, Tijana Osum, Kevin C. Ruggiero, Jenna L. Rust, Blake J. Landry, Samuel J. Jenkins, Marc K. McLachlan, James B. Fife, Brian T. Nat Commun Article 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. Nature Publishing Group 2016-06-13 /pmc/articles/PMC4909947/ /pubmed/27292946 http://dx.doi.org/10.1038/ncomms11804 Text en Copyright © 2016, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Spanier, Justin A.
Frederick, Daniel R.
Taylor, Justin J.
Heffernan, James R.
Kotov, Dmitri I.
Martinov, Tijana
Osum, Kevin C.
Ruggiero, Jenna L.
Rust, Blake J.
Landry, Samuel J.
Jenkins, Marc K.
McLachlan, James B.
Fife, Brian T.
Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment
title Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment
title_full Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment
title_fullStr Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment
title_full_unstemmed Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment
title_short Efficient generation of monoclonal antibodies against peptide in the context of MHCII using magnetic enrichment
title_sort efficient generation of monoclonal antibodies against peptide in the context of mhcii using magnetic enrichment
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4909947/
https://www.ncbi.nlm.nih.gov/pubmed/27292946
http://dx.doi.org/10.1038/ncomms11804
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