Arginine derivative wash in protein purification using affinity chromatography
First Claim
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1. A method for isolating a product, the method comprising:
- (a) providing a load fluid comprising a product and one or more impurities, wherein the product is an Fc-containing monoclonal antibody;
(b) contacting the load fluid with a medium, wherein the medium is a Protein A chromatography column, and wherein the medium can bind the product under conditions suitable for binding the product, thereby providing a bound medium;
(c) contacting the bound medium with one or more wash solutions, wherein at least one wash solution comprises an arginine derivative, thereby providing a washed medium, wherein the arginine derivative is acetyl arginine, agmatine, arginic acid, N-alpha-butyroyl-L-arginine or N-alpha-pyvaloyl arginine;
(d) contacting the washed medium with an elution solution under conditions suitable for eluting the product, and(e) collecting an eluate comprising the product.
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Abstract
The invention relates to methods for isolating a product and/or reducing turbidity and/or impurities from a load fluid comprising the product and one or more impurities by passing the load fluid through a medium, followed by at least one wash solution comprising an arginine derivative, and collecting the product using an elution solution. The invention further relates to a product prepared using a method as described herein.
115 Citations
47 Claims
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1. A method for isolating a product, the method comprising:
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(a) providing a load fluid comprising a product and one or more impurities, wherein the product is an Fc-containing monoclonal antibody; (b) contacting the load fluid with a medium, wherein the medium is a Protein A chromatography column, and wherein the medium can bind the product under conditions suitable for binding the product, thereby providing a bound medium; (c) contacting the bound medium with one or more wash solutions, wherein at least one wash solution comprises an arginine derivative, thereby providing a washed medium, wherein the arginine derivative is acetyl arginine, agmatine, arginic acid, N-alpha-butyroyl-L-arginine or N-alpha-pyvaloyl arginine; (d) contacting the washed medium with an elution solution under conditions suitable for eluting the product, and (e) collecting an eluate comprising the product. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 33)
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26. A method for isolating an antibody, the method comprising:
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(a) providing a load fluid comprising the antibody and one or more impurities, wherein the antibody is an Fc-containing monoclonal antibody; (b) contacting the load fluid with a Protein A medium, wherein the medium can bind the antibody under conditions suitable for binding the antibody, thereby providing a bound medium; (c) contacting the bound medium with one or more wash solutions, wherein at least one wash solution comprises an arginine derivative in a concentration of about 0.1 M to about 2.0 M, thereby providing a washed medium, wherein the arginine derivative is acetyl arginine, agmatine, arginic acid, N-alpha-butyroyl-L-arginine or N-alpha-pyvaloyl arginine; (d) contacting the washed medium with an elution solution under conditions suitable for eluting the antibody; and (e) collecting an eluate comprising the antibody, wherein the ratio of the antibody to host cell protein in the eluate is increased and wherein the eluate has reduced turbidity compared to an eluate recovered in a corresponding method in which no detectable amount of arginine derivative is used in a wash solution. - View Dependent Claims (27, 28, 29, 30, 31, 32)
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34. A method for reducing turbidity in an eluate comprising a product, the method comprising:
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(a) providing a load fluid comprising the product and one or more impurities, wherein the product is an Fc-containing monoclonal antibody; (b) contacting the load fluid with a medium, wherein the medium is a Protein A chromatography column, and wherein the medium can bind the product under conditions suitable for binding the product, thereby providing a bound medium; (c) contacting the bound medium with one or more wash solutions, wherein at least one wash solution comprises an arginine derivative, thereby providing a washed medium, wherein the arginine derivative is acetyl arginine, agmatine, arginic acid, N-alpha-butyroyl-L-arginine or N-alpha-pyvaloyl arginine; (d) contacting the washed medium with an elution solution under conditions suitable for eluting the product, thereby generating an eluate comprising the product; and (e) adjusting the pH of the eluate to between about 6.5 and about 8.2, wherein the eluate has reduced turbidity compared to a corresponding method in which less than about 0.1M of arginine derivative is used in a wash solution. - View Dependent Claims (35, 36, 37, 38, 39, 40, 41)
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42. A method for reducing turbidity and impurities in an eluate comprising a product, the method comprising:
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(a) providing a load fluid comprising the product and one or more impurities, wherein the product is an Fc-containing monoclonal antibody; (b) contacting the load fluid with a medium, wherein the medium is a Protein A chromatography column, and wherein the medium can bind the product under conditions suitable for binding the product, thereby providing a bound medium; (c) contacting the bound medium with one or more wash solutions, wherein at least one wash solution comprises an arginine derivative, thereby providing a washed medium, wherein the arginine derivative is acetyl arginine, agmatine, arginic acid, N-alpha-butyroyl-L-arginine or N-alpha-pyvaloyl arginine; (d) contacting the washed medium with an elution solution under conditions suitable for eluting the product, thereby generating an eluate comprising the product; and (e) adjusting the pH of the eluate to between about 6.5 and about 8.2, wherein the ratio of the product to at least one impurity in the eluate is increased and the eluate has reduced turbidity compared to a corresponding method in which less than about 0.1M arginine derivative is used in a wash solution. - View Dependent Claims (43, 44, 45, 46, 47)
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Specification