TY - JOUR
T1 - Cell surface display of functional human MHC class II proteins
T2 - Yeast display versus insect cell display
AU - Wen, Fei
AU - Sethi, Dhruv K.
AU - Wucherpfennig, Kai W.
AU - Zhao, Huimin
N1 - Funding Information:
This work was supported by the Department of Chemical and Biomolecular Engineering of the University of Illinois at Urbana-Champaign and grants from the National Institutes of Health to K.W.W. (PO1 AI045757, R01AI054520).
PY - 2011/9
Y1 - 2011/9
N2 - Reliable and robust systems for engineering functional major histocompatibility complex class II (MHCII) proteins have proved elusive. Availability of such systems would enable the engineering of peptide-MHCII (pMHCII) complexes for therapeutic and diagnostic applications. In this paper, we have developed a system based on insect cell surface display that allows functional expression of heterodimeric DR2 molecules with or without a covalently bound human myelin basic protein (MBP) peptide, which is amenable to directed evolution of DR2-MBP variants with improved T cell receptor (TCR)-binding affinity. This study represents the first example of functional display of human pMHCII complexes on insect cell surface. In the process of developing this pMHCII engineering system, we have also explored the potential of using yeast surface display for the same application. Our data suggest that yeast display is a useful system for analysis and engineering of peptide binding of MHCII proteins, but not suitable for directed evolution of pMHC complexes that bind with low affinity to self-reactive TCRs.
AB - Reliable and robust systems for engineering functional major histocompatibility complex class II (MHCII) proteins have proved elusive. Availability of such systems would enable the engineering of peptide-MHCII (pMHCII) complexes for therapeutic and diagnostic applications. In this paper, we have developed a system based on insect cell surface display that allows functional expression of heterodimeric DR2 molecules with or without a covalently bound human myelin basic protein (MBP) peptide, which is amenable to directed evolution of DR2-MBP variants with improved T cell receptor (TCR)-binding affinity. This study represents the first example of functional display of human pMHCII complexes on insect cell surface. In the process of developing this pMHCII engineering system, we have also explored the potential of using yeast surface display for the same application. Our data suggest that yeast display is a useful system for analysis and engineering of peptide binding of MHCII proteins, but not suitable for directed evolution of pMHC complexes that bind with low affinity to self-reactive TCRs.
KW - directed evolution
KW - DR2-myelin basic protein (MBP)
KW - insect cell display
KW - major histocompatibility complex (MHC)
KW - yeast display
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U2 - 10.1093/protein/gzr035
DO - 10.1093/protein/gzr035
M3 - Article
C2 - 21752831
AN - SCOPUS:80052325154
SN - 1741-0126
VL - 24
SP - 701
EP - 709
JO - Protein Engineering, Design and Selection
JF - Protein Engineering, Design and Selection
IS - 9
ER -