Engineering fibronectin-based binding proteins by yeast surface display

Tiffany F. Chen; Seymour De Picciotto; Benjamin J. Hackel; K. Dane Wittrup

doi:10.1016/B978-0-12-394292-0.00014-X

Engineering fibronectin-based binding proteins by yeast surface display

Tiffany F. Chen, Seymour De Picciotto, Benjamin J. Hackel, K. Dane Wittrup

Research output: Chapter in Book/Report/Conference proceeding › Chapter

49 Scopus citations

Abstract

Yeast surface display (YSD) presents proteins on the surface of yeast through interaction of the agglutinin subunits Aga1p and Aga2p. The human 10th type III fibronectin (Fn3) is a small, 10-kDa protein domain that maintains its native fold without disulfide bonds. A YSD library of Fn3s has been engineered with a loop amino acid composition similar to that of human antibody complementarity-determining region heavy chain loop 3 (CDR-H3) and varying loop lengths, which has been shown to improve binding ability. There are many advantages of using these small, stable domains that maintain binding capabilities similar to that of antibodies. Here, we outline a YSD methodology to isolate Fn3 binders to a diverse set of target antigens.

Original language	English (US)
Title of host publication	Methods in Protein Design
Publisher	Academic Press Inc.
Pages	303-326
Number of pages	24
ISBN (Print)	9780123942920
DOIs	https://doi.org/10.1016/B978-0-12-394292-0.00014-X
State	Published - 2013

Publication series

Name	Methods in Enzymology
Volume	523
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Bibliographical note

Funding Information:
Funding sources are from the Sanofi-aventis Biomedical Innovation award program, the NIH/NIGMS Biotechnology Training program, the Gordon & Adele Binder Fellowship, NIH Transformative R01 Program (R01EB010246-02), and Integrative Cancer Biology Program (ICBP) at MIT.

Keywords

Alternative scaffolds
Directed evolution
Fibronectin type III domain
Protein engineering
Yeast surface display

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10.1016/B978-0-12-394292-0.00014-X

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title = "Engineering fibronectin-based binding proteins by yeast surface display",

abstract = "Yeast surface display (YSD) presents proteins on the surface of yeast through interaction of the agglutinin subunits Aga1p and Aga2p. The human 10th type III fibronectin (Fn3) is a small, 10-kDa protein domain that maintains its native fold without disulfide bonds. A YSD library of Fn3s has been engineered with a loop amino acid composition similar to that of human antibody complementarity-determining region heavy chain loop 3 (CDR-H3) and varying loop lengths, which has been shown to improve binding ability. There are many advantages of using these small, stable domains that maintain binding capabilities similar to that of antibodies. Here, we outline a YSD methodology to isolate Fn3 binders to a diverse set of target antigens.",

keywords = "Alternative scaffolds, Directed evolution, Fibronectin type III domain, Protein engineering, Yeast surface display",

author = "Chen, {Tiffany F.} and {De Picciotto}, Seymour and Hackel, {Benjamin J.} and Wittrup, {K. Dane}",

note = "Funding Information: Funding sources are from the Sanofi-aventis Biomedical Innovation award program, the NIH/NIGMS Biotechnology Training program, the Gordon & Adele Binder Fellowship, NIH Transformative R01 Program (R01EB010246-02), and Integrative Cancer Biology Program (ICBP) at MIT.",

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AU - Chen, Tiffany F.

AU - De Picciotto, Seymour

AU - Hackel, Benjamin J.

AU - Wittrup, K. Dane

N1 - Funding Information: Funding sources are from the Sanofi-aventis Biomedical Innovation award program, the NIH/NIGMS Biotechnology Training program, the Gordon & Adele Binder Fellowship, NIH Transformative R01 Program (R01EB010246-02), and Integrative Cancer Biology Program (ICBP) at MIT.

PY - 2013

Y1 - 2013

N2 - Yeast surface display (YSD) presents proteins on the surface of yeast through interaction of the agglutinin subunits Aga1p and Aga2p. The human 10th type III fibronectin (Fn3) is a small, 10-kDa protein domain that maintains its native fold without disulfide bonds. A YSD library of Fn3s has been engineered with a loop amino acid composition similar to that of human antibody complementarity-determining region heavy chain loop 3 (CDR-H3) and varying loop lengths, which has been shown to improve binding ability. There are many advantages of using these small, stable domains that maintain binding capabilities similar to that of antibodies. Here, we outline a YSD methodology to isolate Fn3 binders to a diverse set of target antigens.

AB - Yeast surface display (YSD) presents proteins on the surface of yeast through interaction of the agglutinin subunits Aga1p and Aga2p. The human 10th type III fibronectin (Fn3) is a small, 10-kDa protein domain that maintains its native fold without disulfide bonds. A YSD library of Fn3s has been engineered with a loop amino acid composition similar to that of human antibody complementarity-determining region heavy chain loop 3 (CDR-H3) and varying loop lengths, which has been shown to improve binding ability. There are many advantages of using these small, stable domains that maintain binding capabilities similar to that of antibodies. Here, we outline a YSD methodology to isolate Fn3 binders to a diverse set of target antigens.

KW - Alternative scaffolds

KW - Directed evolution

KW - Fibronectin type III domain

KW - Protein engineering

KW - Yeast surface display

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SN - 9780123942920

T3 - Methods in Enzymology

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BT - Methods in Protein Design

PB - Academic Press Inc.

ER -

Engineering fibronectin-based binding proteins by yeast surface display

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