Refolding method
OwnerAlan Roy Fersht, Myriam Marlenne Altamiranoinventors
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Abstract
The invention relates to a method for promoting the folding of a polypeptide, comprising the step of contacting the polypeptide with a molecular chaperone and a foldase.
Claims34
A method for promoting the folding of a polypeptide which method comprises contacting the polypeptide with a molecular chaperone and a foldase, wherein the foldase and optionally the molecular chaperone are immobilised onto a solid phase support.
A solid phase support having immobilised thereon a foldase.
A solid phase support having immobilised thereon a molecular chaperone and a foldase.
A method for immobilising a disulphide-containing peptide onto a solid phase support, comprising the steps of:
- a) reducing the disulphide in the polypeptide with a reducing agent, and removing the reducing agent under conditions so as to prevent re-oxidation;
- b) reversibly blocking the thiol groups of the polypeptide;
- c) contacting the solid phase with the thiol-blocked polypeptide at a non-acidic pH;
- d) blocking any remaining active groups and removing uncoupled polypeptide by washing;
- and
- e) regenerating the thiol groups on the bound polypeptide.
Use of a molecular chaperone and a foldase for promoting the folding of a polypeptide wherein the foldase and optionally the molecular chaperone are immobilised on a solid phase support.
A composition comprising a combination of a molecular chaperone and a foldase wherein the molecular chaperone and the foldase are immobilised on a solid phase support.
A method for promoting the folding of a polypeptide which method comprises contacting the polypeptide with a molecular chaperone and a foldase wherein the molecular chaperone consists essentially of at least one of fragments 191-376, 191-345 and 191-335 of the sequence of intact GroEL, or a homologue thereof.
A composition comprising a combination of a molecular chaperone and a foldase wherein the molecular chaperone consists essentially of at least one of fragments 191-376, 191-345 and 191-335 of the sequence of intact GroEL, or a homologue thereof.
A polypeptide obtained by the method of the invention said polypeptide having greater than 100% of the biological activity of the corresponding native protein.
Dependencies are inferred from the claim text.