Renaturing inclusion bodies

coulier at marseille.inserm.fr coulier at marseille.inserm.fr
Fri Jul 17 01:12:23 EST 1998


Here is my experience, if it can help. We were expressing a protein
(fibroblast growth factor 6, FGF6, with 2 cysteine residues) in E. coli. Most
(maybe 90 or 95 %) of the protein was found in the inclusions bodies, so we
struggled to denature/renature these inclusions bodies. We eventually
succeeded in getting an active growth factor. The yield was something like
500 microgram per liter of culture. We used huge quantities of guanidium and
DTT for the solubilisation, and tedious dialysis steps for the renaturation,
followed by affinity purification on a FPLC column. Then a post-doc came to
the lab, and tried to pass the 5 or 10 % soluble material directly on the
FPLC column (discarding 90-95 % of the protein in the inclusions bodies). No
more guanidium nor DTT, no dialysis, and the yield was around 1-2 mg of
protein per liter of culture.....

François
_______________________________________________________________________
François Coulier,
Institut de Cancérologie et d'Immunologie de Marseille
INSERM Unité 119,
27 bd Leï Roure, 13009 Marseille, France
Tel: 33 (0) 4 91 75 84 11, Fax: 33 (0) 4 91 26 03 64
Mél: coulier at marseille.inserm.fr
http://garlaban.marseille.inserm.fr/u119/home.html

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