High-throughput engineering and analysis of peptide binding to class II MHC.

Proceedings of the National Academy of Sciences of the United States of America
Wei Jiang, Eric T Boder

Abstract

Class II major histocompatibility complex (MHC-II) proteins govern stimulation of adaptive immunity by presenting antigenic peptides to CD4+ T lymphocytes. Many allelic variants of MHC-II exist with implications in peptide presentation and immunity; thus, high-throughput experimental tools for rapid and quantitative analysis of peptide binding to MHC-II are needed. Here, we present an expression system wherein peptide and MHC-II are codisplayed on the surface of yeast in an intracellular association-dependent manner and assayed by flow cytometry. Accordingly, the relative binding of different peptides and/or MHC-II variants can be assayed by genetically manipulating either partner, enabling the application of directed evolution approaches for high-throughput characterization or engineering. We demonstrate the application of this tool to map the side-chain preference for peptides binding to HLA-DR1 and to evolve novel HLA-DR1 mutants with altered peptide-binding specificity.

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Citations

Feb 16, 2012·Nucleic Acids Research·Kyle JacobyAndrew M Scharenberg
Jul 15, 2011·Protein Engineering, Design & Selection : PEDS·Fei WenHuimin Zhao
Aug 7, 2013·Journal of Structural Biology·Christian ReichenAndreas Plückthun
Jul 26, 2011·Journal of Immunological Methods·Guang Lan ZhangVladimir Brusic
Oct 11, 2012·Immunological Reviews·Michael E BirnbaumK Christopher Garcia
May 27, 2014·Cell·Michael E BirnbaumK Christopher Garcia
Dec 25, 2017·Applied Microbiology and Biotechnology·Zhenlin Han, Wei Wen Su
Aug 10, 2018·Frontiers in Immunology·David Gfeller, Michal Bassani-Sternberg
Feb 11, 2020·Frontiers in Bioengineering and Biotechnology·Meng MeiLi Yi
Dec 29, 2017·The Journal of Biological Chemistry·John H McGeeGregory L Verdine
Dec 6, 2019·MedChemComm·Sara LincianoAlessandro Angelini
Jun 13, 2021·Cellular & Molecular Immunology·Rongzeng LiuElizabeth D Mellins

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