title: The Optimization of Bioorthogonal Epitope Ligation within MHC-I Complexes.
abstract: Antigen recognition followed by the activation of cytotoxic T-cells (CTLs) is a key step in adaptive immunity, resulting in clearance of viruses and cancers. The repertoire of peptides that have the ability to bind to the major histocompatibility type-I (MHC-I) is enormous, but the approaches available for studying the diversity of the peptide repertoire on a cell are limited. Here, we explore the use of bioorthogonal chemistry to quantify specific peptide-MHC-I complexes (pMHC-I) on cells. We show that modifying epitope peptides with bioorthogonal groups in surface accessible positions allows wild-type-like MHC-I binding and bioorthogonal ligation using fluorogenic chromophores in combination with a Cu(I)-catalyzed Huisgen cycloaddition reaction. We expect that this approach will make a powerful addition to the antigen presentation toolkit as for the first time it allows quantification of antigenic peptides for which no detection tools exist.
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Joanna B
authors.initials: JB
authors.lastname: Pawlak
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center and The Institute for Chemical Immunology , P.O. Box 9600, 2300 RC Leiden, The Netherlands.
authors.forename: Brett J
authors.initials: BJ
authors.lastname: Hos
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Michel J
authors.initials: MJ
authors.lastname: van de Graaff
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Otty A
authors.initials: OA
authors.lastname: Megantari
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Nico
authors.initials: N
authors.lastname: Meeuwenoord
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Filippov
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center and The Institute for Chemical Immunology , P.O. Box 9600, 2300 RC Leiden, The Netherlands.
authors.forename: Ferry
authors.initials: F
authors.lastname: Ossendorp
authors.affiliation: Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University Einsteinweg 55, 2333 CC Leiden, The Netherlands.
authors.forename: Sander I
authors.initials: SI
authors.lastname: van Kasteren
mesh_terms: D000939:Epitopes / Q000737:chemistry*; D015395:Histocompatibility Antigens Class I / Q000737:chemistry*; D006801:Humans; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D000939:Epitopes; D015395:Histocompatibility Antigens Class I