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.
| Original language | English (US) |
|---|---|
| Article number | 11804 |
| Journal | Nature communications |
| Volume | 7 |
| DOIs | |
| State | Published - Jun 13 2016 |
Bibliographical note
Funding Information:This work was supported by NIH R01 AI106791 (B.T.F.), NIH U24-AI118635 (B.T.F.), NIH P01 AI35296 (B.T.F. and M.K.J.), NIH R01 AI103343 (J.B.M.), Irvington Fellowship Program of the Cancer Research Institute (J.J.T.), Juvenile Diabetes Research Foundation 2-2011-662 (B.T.F.) and 3-2014-215 (J.A.S.).
