Methods of purifying anti A β antibodies
First Claim
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1. A method for purifying an Aβ
- binding protein having an Fc region from a source liquid comprising the protein and one or more impurities, wherein the Aβ
binding protein is an anti-Aβ
antibody, comprising the steps of;
adsorbing the Aβ
binding protein to an Fc binding agent;
washing the Fc binding agent bound to the Aβ
binding protein with a buffer solution containing CaCl2 at a concentration from about 0.5 M to about 3 M to reduce the one or more impurities; and
recovering the Aβ
binding protein from the Fc binding agent in an elution solution.
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Abstract
The present application provides methods of purifying Aβ binding proteins having a Fc region, for example, anti-Aβ antibodies or antibody fusions, by adsorbing the Aβ binding protein to a Fc binding agent, such as, for example, Protein A or Protein G, followed by a wash with a divalent cation salt buffer to remove impurities and subsequent recovery of the adsorbed Aβ binding protein. The present application also features methods of eluting the purified Aβ binding protein as well as the incorporation of the methods within a purification train. Kits comprising components for carrying out the methods and instructions for use are also provided.
97 Citations
96 Claims
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1. A method for purifying an Aβ
- binding protein having an Fc region from a source liquid comprising the protein and one or more impurities, wherein the Aβ
binding protein is an anti-Aβ
antibody, comprising the steps of;adsorbing the Aβ
binding protein to an Fc binding agent;washing the Fc binding agent bound to the Aβ
binding protein with a buffer solution containing CaCl2 at a concentration from about 0.5 M to about 3 M to reduce the one or more impurities; andrecovering the Aβ
binding protein from the Fc binding agent in an elution solution. - 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, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
- binding protein having an Fc region from a source liquid comprising the protein and one or more impurities, wherein the Aβ
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41. A method for purifying an Aβ
- binding protein having an Fc region from a source liquid comprising the Aβ
binding protein and one or more impurities, wherein the Aβ
binding protein is a humanized 3D6 antibody and wherein the one or more impurities comprise one or more intron read through variant species (IRT), the method comprising the steps of;adsorbing the Aβ
binding protein to an affinity ligand which is an Fc binding agent;washing the affinity ligand with a buffer solution containing CaCl2 at a concentration from about 0.5 M to about 3 M to reduce the IRT; and recovering the Aβ
binding protein from the affinity ligand in an elution solution, wherein the Aβ
binding protein having an Fc region is purified from the source liquid. - View Dependent Claims (42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65)
- binding protein having an Fc region from a source liquid comprising the Aβ
-
66. A method for purifying an Aβ
- binding protein having an Fc region from a source liquid comprising the protein and one or more impurities, wherein the Aβ
binding protein is a humanized anti-Aβ
antibody, the method comprising the steps of;adsorbing the Aβ
binding protein to an affinity ligand which is an Fc binding agent;washing the affinity ligand with a buffer solution containing CaCl2 at a concentration from about 0.5 M to about 3 M to reduce the one or more impurities; and recovering the Aβ
binding protein from the affinity ligand in an elution solution. - View Dependent Claims (67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95)
- binding protein having an Fc region from a source liquid comprising the protein and one or more impurities, wherein the Aβ
-
96. A method for purifying an Aβ
- binding protein having an Fc region from a source liquid comprising the Aβ
binding protein and one or more impurities, the method comprising the steps of;adsorbing the Aβ
binding protein to an Fc binding agent comprising Protein A, wherein the Aβ
binding protein is a humanized 3D6 antibody comprising a light chain comprising the amino acid sequence of SEQ ID NO;
1 and a heavy chain comprising the amino acid sequence of residues 1-448 of SEQ ID NO;
2;washing the Fc binding agent bound to the Aβ
binding protein with a buffer solution containing CaCl2 at a concentration of from about 1.5 M to about 2 M to reduce the one or more impurities; andrecovering the Aβ
binding protein from the Fc binding agent using an elution buffer having a pH in a range from about 2.5 to about 3.5,wherein the Aβ
binding protein having an Fc region is purified from the source liquid.
- binding protein having an Fc region from a source liquid comprising the Aβ
Specification