Chapter title |
Creation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain
|
---|---|
Chapter number | 13 |
Book title |
SH2 Domains
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6762-9_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6760-5, 978-1-4939-6762-9
|
Authors |
Haiming Huang, Tomonori Kaneko, Sachdev S. Sidhu, Shawn S. C. Li, Huang, Haiming, Kaneko, Tomonori, Sidhu, Sachdev S., Li, Shawn S. C. |
Editors |
Kazuya Machida, Bernard A. Liu |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 14 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 3 | 21% |
Student > Bachelor | 2 | 14% |
Researcher | 2 | 14% |
Other | 1 | 7% |
Professor > Associate Professor | 1 | 7% |
Other | 0 | 0% |
Unknown | 5 | 36% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 4 | 29% |
Chemistry | 2 | 14% |
Agricultural and Biological Sciences | 1 | 7% |
Computer Science | 1 | 7% |
Unknown | 6 | 43% |