REFOLDING CHROMATOGRAPHY WITH IMMOBILIZED MINI-CHAPERONES

Mini-chaperones (e.g., a peptide consisting of residues 191-345 of Gro EL) that are immobilized on agarose have very efficient chaperoning ac tivity with several proteins that are otherwise recalcitrant to renatu ration by conventional methods. We have used immobilized mini-chaperon es both in column chromatography and batchwise to renature an insolubl e protein from an inclusion body, to refold apparently irreversibly de natured proteins, and to recondition enzymes that have lost activity o n storage. Refolding chromatography offers an efficient and simple mea ns to renature proteins in high yield and with biological activity.