title: Preparation of stable single-chain trimers engineered with peptide, beta2 microglobulin, and MHC heavy chain.
abstract: This unit describes a method for constructing a class I MHC molecule with a bound peptide as a single polypeptide chain, termed SCT, for single chain trimer. The component organization of the SCT appears to be widely applicable to different mouse or human MHC class I isotypes bound by different antigenic peptides. The enhanced peptide occupancy afforded by the SCT format makes these molecules effective reagents as DNA vaccines, multimeric staining reagents to enumerate CD8 T cells, and probes of lymphocyte biology.
authors.affiliation: Washington University School of Medicine, St. Louis, Missouri.
authors.forename: Ted
authors.initials: T
authors.lastname: Hansen
authors.affiliation: Washington University School of Medicine, St. Louis, Missouri.
authors.forename: Y Y Lawrence
authors.initials: YYL
authors.lastname: Yu
authors.affiliation: Washington University School of Medicine, St. Louis, Missouri.
authors.forename: Daved H
authors.initials: DH
authors.lastname: Fremont
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002850:Chromatography, Gel; D004220:Disulfides / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006183:H-2 Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015234:HLA-A Antigens / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D014162:Transfection; D001613:beta 2-Microglobulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004220:Disulfides; D006183:H-2 Antigens; D015234:HLA-A Antigens; D015395:Histocompatibility Antigens Class I; D010455:Peptides; D011994:Recombinant Proteins; D001613:beta 2-Microglobulin