Stimulus responsive polymers for the purification of biomolecules
AssigneeEMD Millipore Corporation
InventorsJad Jaber, Wilson Moya, James Hamzik, Arezki Boudif, Yu Zhang, Neil Soice, John Charkoudian, Nripen Singh
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Abstract
The present invention provides novel and improved stimulus responsive polymers and methods of using the same for the purification of biomolecules.
Claims41
A soluble stimulus responsive polymer comprising a polyvinylamine backbone comprising one or more hydrophobic groups attached to the backbone, wherein the polymer is capable of binding and precipitating a biomolecule of interest in a sample upon addition of a stimulus.
A method of separating a target molecule from one or more impurities in a sample, wherein the method comprises the steps of:
- (a) providing a sample comprising a target molecule and one or more impurities;
- (b) contacting the sample with a soluble stimulus responsive polymer comprising a polyelectrolyte backbone comprising one or more hydrophobic groups attached to the backbone thereby to form a complex of polymer and the one or more impurities;
- and
- (c) adding a stimulus to the sample, thereby to precipitate the complex out of solution,
- thereby to separate the target molecule from one or more impurities.
A soluble stimulus responsive polymer comprising the following structure:
- Wherein the polymer comprises a carbon containing polyelectrolyte backbone;
- R1, R2 and R3 are charged groups which form a part of the backbone;
- R4 is a hydrophobic group attached to a charged group in the backbone;
- R5 is a group possessing a charge opposite of the charge found in R1, R2 or R3;
- and n is the number of monomeric units in the polymer, wherein n is equal to or greater than 2.
A method of separating an antibody from one or more impurities in a sample while minimizing residual amounts of polymer, the method comprising the steps of:
- (a) providing a sample comprising an antibody and one or more impurities;
- (b) contacting the sample with a soluble stimulus responsive polymer comprising a polyelectrolyte backbone comprising one or more hydrophobic groups attached to the backbone under a first set of conditions suitable for the polymer to bind one or more impurities, thereby to form a first complex of polymer and one or more impurities, wherein the first set of conditions comprise adjusting pH or salt concentration of the sample during or after the addition of the polymer;
- (c) precipitating the first complex from the sample under a second set of conditions;
- (d) contacting the sample with a stimulus comprising a multivalent ion, thereby to form a second complex of residual polymer and multivalent ion;
- and
- (e) precipitating the second complex;
- thereby to separate the antibody from one or more impurities in the sample while reducing the amount of residual polymer in the sample.
A soluble stimulus responsive polymer for separating a target molecule from one or more impurities comprising the following formula:
- wherein the polymer comprises a carbon containing polyelectrolyte backbone;
- R1, R2 and R3 are amine groups which form a part of the backbone;
- R4 is a phenyl group which is attached to 5% to 75% of the R1, R2 or R3 groups;
- R5 is a cation exchange group modifying 5% to 50% of the R1, R2 or R3 groups;
- and n is the number of monomeric units in the polymer, wherein n is equal to or greater than 2.
A method of separating an antibody from one or more impurities in a sample using a stimulus responsive polymer, while minimizing residual amounts of polymer, the method comprising the steps of:
- (a) providing a sample comprising an antibody and one or more impurities;
- (b) contacting the sample with a soluble stimulus responsive polymer comprising a polyelectrolyte backbone comprising one or more hydrophobic groups attached to the backbone under a first set of conditions suitable for the polymer to bind one or more impurities, thereby to form a first complex of polymer and one or more impurities, wherein the first set of conditions comprise adjusting pH or salt concentration of the sample during or after the addition of the polymer;
- (c) flocculating the first complex from the sample under a first set of conditions;
- (d) removing the flocculated solids;
- (e) contacting the sample with a multivalent ion, thereby to form a second complex of residual polymer and multivalent ion;
- and
- (f) precipitating the second complex
- thereby to separate the antibody from one or more impurities in the sample while reducing the amount of residual polymer in the sample.
A method of separating a target molecule from one or more impurities in a sample, wherein the method comprises the steps of:
- (a) providing a sample comprising a target molecule and one or more impurities;
- (b) contacting the sample with a soluble multivalent ion responsive polymer comprising a polyelectrolyte backbone comprising one or more hydrophobic groups attached to the backbone, wherein a multivalent ion is added during or after addition of the polymer, thereby to form a complex of polymer and the one or more impurities;
- (c) precipitating the complex out of solution,
- thereby to separate the target molecule from one or more impurities.
Dependencies are inferred from the claim text.