Departmental Papers (CBE)

Document Type

Journal Article

Date of this Version

November 2005

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 wildtype MHCII. Results further point to the potential for using yeast surface display for engineering and analyzing the antigen binding properties of MHCII.

Comments

Postprint version. “This is a preprint of an article published in Biotechnology and Bioengineering, Volume 92, Issue 4, November 20, 2005, pages 485-491.”
Publisher URL: http://dx.doi.org/10.1002/bit.20616

Keywords

yeast display, class II MHC, peptide loading, heterodimer expression in yeast, protein engineering

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Date Posted: 24 March 2006

This document has been peer reviewed.