High-Throughput Engineering and Analysis of Class II Mhc/Peptide Binding by Yeast Co-Display

Loading...
Thumbnail Image
Degree type
Doctor of Philosophy (PhD)
Graduate group
Chemical and Biomolecular Engineering
Discipline
Subject
yeast surface display
MHC-II
directed evolution
peptide binding
yeast co-display
Biochemical and Biomolecular Engineering
Funder
Grant number
License
Copyright date
Distributor
Related resources
Contributor
Abstract

Polymorphisms of major histocompatibility complex (MHC) and molecular mechanisms of their antigen-presenting specificity and promiscuity have great impact on T cell-mediated immune responses and related diseases. Challenges in elucidating the characteristics of antigenic peptide binding by MHC motivate the development of high throughput experimental tools to quantitatively analyze interactions between hundreds of MHC allelic proteins and various peptide sequences. We demonstrated such a method by co-displaying target peptides and class II MHC (MHC-II) on the yeast surface in an intracellular association-dependent manner. The optimized yeast co-display system enabled quantitative mapping of side-chain preferences and general motifs for peptides binding to MHC-II by site-directed mutagenesis or peptide library screening, and also allowed rapid tailoring of MHC-II peptide binding specificity by directed evolution approaches, which derived MHC-II allelic mutants with altered peptide binding specificity or hyper-promiscuity. Comparison of these experimentally engineered mutants with naturally discovered MHC-II proteins recovered valuable information about structure-function relationship in the evolutionary mechanisms for polymorphic MHC-II molecules, which could direct future immunotherapeutic innovation.

Advisor
Wei Jiang
Date of degree
2010-05-17
Date Range for Data Collection (Start Date)
Date Range for Data Collection (End Date)
Digital Object Identifier
Series name and number
Volume number
Issue number
Publisher
Publisher DOI
Journal Issue
Comments
Recommended citation