Larsson, Anna
- Aix-Marseille Université
Sammanfattning
Producing recombinant proteins in Escherichia coli (E. coli) is generally performed using a trial and error approach with the different expression variables being tested independently from each other. As a consequence, variable interactions are lost which makes the trial and error approach quite time-consuming. In this paper, we report how switching from a trial and error to a fractional factorial approach allows testing in less than 2 weeks four expression variables (E. coli strains, culture media, expression temperatures and N-terminal fusion tags) in a single experiment. The method, called "Fusion-InFFact", was validated using four test proteins. In all cases, Fusion-InFFact allowed finding conditions for expressing high yields of soluble proteins. The method was originally set-up for high throughput structural genomics programs, but can be used in any recombinant protein expression project. © 2012 Published by Elsevier Inc.
Nyckelord
Design of experiment; Escherichia coli; Fractional factorial approach; Recombinant protein; Structural genomics
Publicerad i
Protein Expression and Purification
2012, volym: 84, nummer: 2, sidor: 204-213
SLU författare
UKÄ forskningsämne
Bioprocessteknik
Publikationens identifierare
- DOI: https://doi.org/10.1016/j.pep.2012.05.011
Permanent länk till denna sida (URI)
https://res.slu.se/id/publ/132279