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Enhancedin vitrosynthesis of active proteins containing disulfide bonds

  • US 7,041,479 B2
  • Filed: 03/31/2003
  • Issued: 05/09/2006
  • Est. Priority Date: 09/06/2000
  • Status: Expired due to Term
First Claim
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1. A method for in vitro synthesis of properly folded polypeptides comprising at least one disulfide bond, in a reaction mix comprising a biological extract comprising components of polypeptide synthesis machinery, wherein such components are capable of expressing a nucleic acid encoding a desired polypeptide, the method comprising:

  • synthesizing said polypeptide in a reaction mix wherein the biological extract has been pre-treated with a sulfhydryl inactivating agent that irreversibly inactivates free sulfhydryl groups;

    isolating said polypeptide from said reaction mixture, wherein the amount of said polypeptide that is properly folded is enhanced relative to polypeptide synthesized in the absence of pretreatment with said sulfhydryl inactivating agent.

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