Refolding proteins using a chemically controlled redox state

  • US 9,856,287 B2
  • Filed: 02/01/2017
  • Issued: 01/02/2018
  • Est. Priority Date: 06/22/2009
  • Status: Active Grant
First Claim
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1. A method of refolding proteins expressed in a non-mammalian expression system, the method comprising:

  • contacting the proteins with a preparation that supports the renaturation of at least one of the proteins to a biologically active form, to form a refold mixture, the preparation comprising;

    at least one ingredient selected from the group consisting of a denaturant, an aggregation suppressor and a protein stabilizer;

    an amount of oxidant; and

    an amount of reductant,wherein the amounts of the oxidant and the reductant are related through a thiol-pair ratio and a thiol-pair buffer strength,wherein the thiol-pair ratio is in the range of 0.001-100; and

    wherein the thiol-pair buffer strength maintains the solubility of the preparation; and

    incubating the refold mixture so that at least about 25% of the proteins are properly refolded.

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