Creation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain.

TitleCreation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain.
Publication TypeJournal Article
Year of Publication2017
AuthorsHuang, Haiming, Kaneko Tomonori, Sidhu Sachdev S., and Li Shawn S. C.
JournalMethods Mol Biol
Volume1555
Pagination225-254
Date Published2017
ISSN1940-6029
Abstract

<p>Commercial antibodies raised against phosphotyrosine have been widely used as reagents to detect or isolate tyrosine-phosphorylated proteins from cellular samples. However, these antibodies are costly and are not amenable to in-house production in an academic lab setting. In this chapter, we describe a method to generate super-high affinity SH2 domains, dubbed the phosphotyrosine superbinders, by evolving a natural SH2 domain using the phage display technology. The superbinders are stable and can be easily produced in Escherichia coli in large quantities. The strategy presented here may also be applied to other protein domains to generate domain variants with markedly enhanced affinities for a specific post-translational modification.</p>

DOI10.1007/978-1-4939-6762-9_13
Alternate JournalMethods Mol. Biol.
PubMed ID28092036

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