Temperature dependent ligand facilitated purification of proteins
AssigneeAffitech AS
InventorsBo Akerström, Tommy Cedervall
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Abstract
The invention concerns the use of a coiled coil protein which is capable of forming a homodimer which dissociates into monomers on change of temperature, in a method of isolating a ligand or desired protein from a sample. A method of isolating a ligand which binds to the coiled coil protein is described as well as methods of isolating a desired protein which is tagged with a coiled coil protein or a ligand which binds to a coiled coil protein, or is tagged with a coiled coil protein monomer.
Claims23
A method of isolating a ligand comprising the steps of:
- a. providing a solid support having bound thereto a coiled coil protein, which binds to said ligand and whose affinity for the ligand is temperature dependent;
- b. contacting a sample containing the ligand with the support at a temperature which promotes binding of the ligand to the protein;
- c. altering the temperature to promote dissociation of the ligand from the protein;
- and
- d. collecting said ligand.
A method of isolating a desired protein comprising the steps of:
- a. tagging said protein with a coiled coil protein monomer;
- b. providing a solid support having bound thereto coiled coil protein monomer;
- c. contacting a sample containing the tagged protein with the support at a temperature which promotes binding between the monomer tagged protein and support-bound monomer;
- d. altering the temperature to promote dissociation of the desired protein-monomer from the support bound monomer;
- and
- e. collecting the desired protein-monomer.
A method of isolating a desired protein comprising the steps of:
- (a) tagging said desired protein with a coiled coil protein whose affinity for a ligand is temperature dependent;
- (b) providing a solid support having bound thereto said ligand;
- (c) contacting a sample containing the tagged protein with the support at a temperature which promotes binding between the coiled coil protein and ligand;
- (d) altering the temperature to promote dissociation of the protein from the ligand;
- and
- (e) collecting said tagged protein.
A method of isolating a desired protein comprising the steps of:
- (a) tagging said desired protein with a ligand which binds to a coiled coil protein whose affinity for a ligand is temperature dependent;
- (b) providing a solid support having bound thereto said coiled coil protein;
- (c) contacting a sample containing the tagged protein with the support at a temperature which promotes binding between the coiled coil protein and ligand;
- (d) altering the temperature to promote dissociation of the tagged protein from the coiled coil protein;
- and
- (e) collecting said tagged protein.
Dependencies are inferred from the claim text.