High-throughput expression screening and purification of recombinant proteins in E-coli

Saez, Natalie J. and Vincentelli, Renaud (2014). High-throughput expression screening and purification of recombinant proteins in E-coli. In Yu Wai Chen (Ed.), Structural genomics: General applications (pp. 33-53) New York United States: Humana Press. doi:10.1007/978-1-62703-691-7_3


Author Saez, Natalie J.
Vincentelli, Renaud
Title of chapter High-throughput expression screening and purification of recombinant proteins in E-coli
Formatted title
High-throughput expression screening and purification of recombinant proteins in E-coli
Title of book Structural genomics: General applications
Place of Publication New York United States
Publisher Humana Press
Publication Year 2014
Sub-type Critical review of research, literature review, critical commentary
DOI 10.1007/978-1-62703-691-7_3
Open Access Status
Year available 2013
Series Methods in Molecular Biology
ISBN 978-1-62703-690-0
978-1-62703-691-7
ISSN 1064-3745
Editor Yu Wai Chen
Volume number 1091
Chapter number 3
Start page 33
End page 53
Total pages 21
Total chapters 24
Collection year 2015
Language eng
Formatted Abstract/Summary
The protocols outlined in this chapter allow for the small-scale test expression of a single or multiple proteins concurrently using several expression conditions to identify optimal strategies for producing soluble, stable proteins. The protocols can be performed manually without the need for specialized equipment, or can be translated to robotic platforms. The high-throughput protocols begin with transformation in a 96-well format, followed by small-scale test expression using auto-induction medium in a 24-well format, finishing with purification in a 96-well format. Even from such a small scale, there is the potential to use the purified proteins for characterization in pilot studies, for sensitive micro-assays, or for the quick detection of and differentiation of the expected size and oxidation state of the protein by mass spectrometry.
Keyword E. coli
Bacteria
Expression
Recombinant
High-throughput
Q-Index Code B1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Book Chapter
Collections: Non HERDC
Institute for Molecular Bioscience - Publications
 
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