Cargando…
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...
Autores principales: | , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1782437918620516352 |
---|---|
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. |
format | Online Article Text |
id | pubmed-4909947 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT spanierjustina efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT frederickdanielr efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT taylorjustinj efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT heffernanjamesr efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT kotovdmitrii efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT martinovtijana efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT osumkevinc efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT ruggierojennal efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT rustblakej efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT landrysamuelj efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT jenkinsmarck efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT mclachlanjamesb efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment AT fifebriant efficientgenerationofmonoclonalantibodiesagainstpeptideinthecontextofmhciiusingmagneticenrichment |