title: Yeast surface display of a noncovalent MHC class II heterodimer complexed with antigenic peptide.
abstract: Microbial protein display technologies have enabled directed molecular evolution of binding and stability properties in numerous protein systems. In particular, dramatic improvements to antibody binding affinity and kinetics have been accomplished using these tools in recent years. Examples of successful application of display technologies to other immunological proteins have been limited to date. Herein, we describe the expression of human class II major histocompatibility complex allele (MHCII) HLA-DR4 on the surface of Saccharomyces cerevisiae as a noncovalently associated heterodimer. The yeast-displayed MHCII is fully native as assessed by binding of conformationally specific monoclonal antibodies; failure of antibodies specific for empty HLA-DR4 to bind yeast-displayed protein indicates antigenic peptide is bound. This report represents the first example of a noncovalent protein dimer displayed on yeast and of successful display of wild-type MHCII. Results further point to the potential for using yeast surface display for engineering and analyzing the antigen binding properties of MHCII.
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA. boder@seas.upenn.edu
authors.forename: Eric T
authors.initials: ET
authors.lastname: Boder
authors.forename: Jerome R
authors.initials: JR
authors.lastname: Bill
authors.forename: Andrew W
authors.initials: AW
authors.lastname: Nields
authors.forename: Philippa C
authors.initials: PC
authors.lastname: Marrack
authors.forename: John W
authors.initials: JW
authors.lastname: Kappler
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D015870:Gene Expression; D015804:HLA-DR4 Antigen / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; D015804:HLA-DR4 Antigen; D011994:Recombinant Proteins