Premium
Efficient solubilization of inclusion bodies
Author(s) -
Freydell Esteban J.,
Ottens Marcel,
Eppink Michel,
van Dedem Gijs,
van der Wielen Luuk
Publication year - 2007
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200700046
Subject(s) - inclusion bodies , solubility , solubilization , monomer , fraction (chemistry) , recombinant dna , chemistry , reagent , escherichia coli , protein aggregation , biochemistry , chromatography , kinetics , organic chemistry , polymer , physics , quantum mechanics , gene
The overexpression of recombinant proteins in Escherichia coli leads in most cases to their accumulation in the form of insoluble aggregates referred to as inclusion bodies (IBs). To obtain an active product, the IBs must be solubilized and thereafter the soluble monomeric protein needs to be refolded. In this work we studied the solubilization behavior of a model‐protein expressed as IBs at high protein concentrations, using a statistically designed experiment to determine which of the process parameters, or their interaction, have the greatest impact on the amount of soluble protein and the fraction of soluble monomer. The experimental methodology employed pointed out an optimum balance between maximum protein solubility and minimum fraction of soluble aggregates. The optimized conditions solubilized the IBs without the formation of insoluble aggregates; moreover, the fraction of soluble monomer was ∼75% while the fraction of soluble aggregates was ∼ 5%. Overall this approach guarantees a better use of the solubilization reagents, which brings an economical and technical benefit, at both large and lab scale and may be broadly applicable for the production of recombinant proteins.