High-throughput protein expression screening and purification in Escherichia coli

Renaud Vincentelli, Agnès Cimino, Arie Geerlof, Atsushi Kubo, Yutaka Satou, Christian Cambillau

Research output: Contribution to journalArticlepeer-review

77 Citations (Scopus)


Escherichia coli (E. coli) is the most widely used expression system for the production of recombinant proteins for structural and functional studies. However, to obtain milligrams of soluble proteins is still challenging since many proteins are expressed in an insoluble form without optimization. Therefore when working with tens of proteins or protein domains it is recommended that high-throughput expression screening at a small scale (1-4. ml of culture) is carried out to identify the optimal conditions for soluble protein production. Once determined, these culture conditions can be applied at a large scale to produce sufficient protein for structural or functional studies. We describe a procedure that has enabled the systematic screening of culture conditions or fusion-tags on hundreds of cultures per week. The analysis of the optimal conditions for the soluble production of these proteins helped us to design a simple and efficient protocol for soluble protein expression screening. This protocol has since been used on hundreds of proteins and is illustrated with the genome wide scale production of proteins containing the DNA binding domains of Ciona intestinalis.

Original languageEnglish
Pages (from-to)65-72
Number of pages8
Issue number1
Publication statusPublished - 2011 Sept


  • Ciona intestinalis
  • Escherichia coli
  • High-throughput
  • Solubility screening
  • Thioredoxin


Dive into the research topics of 'High-throughput protein expression screening and purification in Escherichia coli'. Together they form a unique fingerprint.

Cite this