Refolding Proteins Using a Chemically Controlled Redox State

Abstract

A method of refolding proteins expressed in non-mammalian cells present in concentrations of 2.0 g/L or higher is disclosed. The method comprises identifying foe thiol pair ratio and the redox buffer strength to achieve conditions under which efficient folding at concentrations of 2.0 g/L or higher is achieved and can be employed over a range of volumes, including commercial scale.

1 . A method of refolding a protein expressed in a non-mammalian expression system and present in a volume at a concentration of 2.0 g/L or greater comprising:
 (a) contacting the protein with a refold buffer comprising a redox component comprising a fatal thiol-pair ratio having a range of  0 . 001  to  100  and a redox buffer strength of 2 mM or greater and one or more of:
 (i) a denaturant; 
 (ii) an aggregation suppressor; and 
 (hi) a protein stabilizer; 
   to form a refold mixture;   (b) incubating the refold mixture; and   (c) isolating the protein from the refold mixture.   
     
     
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