US2021284685A1PendingUtilityA1

Methods for removing a contaminant using indigenous protein displacement ion exchange membrane chromatography

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Assignee: GENENTECH INCPriority: Aug 14, 2008Filed: Jan 20, 2021Published: Sep 16, 2021
Est. expiryAug 14, 2028(~2.1 yrs left)· nominal 20-yr term from priority
Inventors:Arick Michael BrownJerome Joseph Bill, Jr.Timothy P. TullyChristopher John Dowd
C07K 1/36C07K 1/18C07K 16/065C07K 1/00C07K 1/34C07K 1/22B01D 15/08
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Claims

Abstract

Methods for purifying a polypeptide from a composition comprising the polypeptide and at least one contaminant are described, which methods comprise the sequential steps of: (a) passing the composition through an ion exchange membrane, where the polypeptide and the membrane have opposite charge, at operating conditions comprised of a buffer having a pH sufficiently distinct from the pI of the polypeptide to enhance the charge of the polypeptide and a low ionic strength effective to prevent the shielding of charges by buffer ions, which cause the membrane to bind the polypeptide and the at least one contaminant, and (b) recovering the purified polypeptide from the effluent.

Claims

exact text as granted — not AI-modified
1 . A method for purifying an antibody from a composition comprising the antibody and at least one contaminant, which method comprises the sequential steps of:
 a. passing the composition through an ion exchange membrane, wherein the antibody and the membrane have opposite charge, at operating conditions comprised of a buffer having a pH sufficiently distinct from the pI of the antibody to enhance the charge of the antibody and a low ionic strength effective to prevent the shielding of charges by buffer ions, which cause the membrane to bind the antibody and the at least one contaminant, and   b. recovering the purified antibody from the effluent.   
     
     
         2 . The method of  claim 1  wherein the ion exchange membrane has a pore size of 0.1 to 100 μm. 
     
     
         3 . A method for purifying an antibody from a composition comprising the antibody and at least one contaminant, which method comprises the sequential steps of:
 a. passing the composition through a cation exchange membrane, wherein the antibody and the membrane have opposite charge, at operating conditions comprised of a buffer having a pH of about 1 to about 5 pH units below the pI of the antibody and a conductivity of < about 40 mS/cm, which cause the membrane to bind the antibody and the at least one contaminant, and   b. recovering the purified antibody from the effluent.   
     
     
         4 . The method of  claim 3  wherein the pH is about 1 to about 4 pH units below the pI of the antibody. 
     
     
         5 . The method of  claim 3  wherein the pH is about 1 to about 3 pH units below the pI of the antibody. 
     
     
         6 . The method of  claim 3  wherein the pH is about 1 to about 2 pH units below the pI of the antibody. 
     
     
         7 . The method of  claim 3  wherein the pH is about 1 pH unit below the pI of the antibody. 
     
     
         8 . The method of  claim 3  wherein the conductivity is < about 20 mS/cm. 
     
     
         9 . The method of  claim 3  wherein the conductivity is < about 10 mS/cm. 
     
     
         10 . A method for purifying an antibody from a composition comprising the antibody and at least one contaminant, which method comprises the sequential steps of:
 a. passing the composition through an anion exchange membrane, wherein the antibody and the membrane have opposite charge, at operating conditions comprised of a buffer having a pH of about 1 to about 5 pH units above the pI of the antibody and a conductivity of < about 40 mS/cm, which cause the membrane to bind the antibody and the at least one contaminant, and   b. recovering the purified antibody from the effluent.   
     
     
         11 . The method of  claim 10  wherein the pH is about 1 to about 4 pH units above the pI of the antibody. 
     
     
         12 . The method of  claim 10  wherein the pH is about 1 to about 3 pH units above the pI of the antibody. 
     
     
         13 . The method of  claim 10  wherein the pH is about 1 to about 2 pH units above the pI of the antibody. 
     
     
         14 . The method of  claim 10  wherein the pH is about 1 pH unit above the pI of the antibody. 
     
     
         15 . The method of  claim 10  wherein the conductivity is < about 20 mS/cm. 
     
     
         16 . The method of  claim 10  wherein the conductivity is < about 10 mS/cm. 
     
     
         17 . The method of  claim 3  wherein the membrane is a mixed mode adsorber. 
     
     
         18 . The method of  claim 3  wherein the contaminant is selected from the group consisting of: leached protein A: nucleic acid: a variant, fragment, aggregate or derivative of the antibody: another antibody, endotoxin: a viral contaminant; and a cell culture media component. 
     
     
         19 . The method of  claim 3  wherein the antibody comprises a CH2/CH3 region. 
     
     
         20 . (canceled) 
     
     
         21 . The method of  claim 1  wherein the antibody is a monoclonal antibody. 
     
     
         22 . The method of  claim 3  further comprising subjecting the composition comprising the antibody to one or more further purification step(s) either before, during, or after steps a through b, said purification step being protein A affinity chromatography. 
     
     
         23 . The method of  claim 3  further comprising subjecting the composition comprising the antibody to one or more further purification step(s) either before, during, or after steps a through b, said purification step being ion exchange chromatography. 
     
     
         24 . The method of  claim 3  further comprising subjecting the composition comprising the antibody to one or more further purification step(s) run continuously during steps a through b, said purification step being ion exchange chromatography. 
     
     
         25 . The method of  claim 3  further comprising preparing a pharmaceutical composition by combining the purified antibody with a pharmaceutically acceptable carrier. 
     
     
         26 . The method of  claim 3  wherein the cation exchange membrane has a pore size of 0.1 to 100 μm. 
     
     
         27 . The method of  claim 3 , wherein the antibody, or antigen-binding fragment thereof, is selected from the group consisting of HER2 antibodies, EGFR antibodies, CD20 antibodies, CD22 antibodies, VEGF antibodies, VEGF receptor antibodies, IgE antibodies, Apo-2 receptor antibodies, and TNF-alpha antibodies. 
     
     
         28 . The method of  claim 27  wherein the antibody is
 (i) a HER2 antibody selected from the group consisting of trastuzumab and pertuzumab; 
 (ii) the CD20 antibody rituximab; 
 (iii) the VEGF antibody bevacizumab; or 
 (iv) the IgE antibody omalizumab.

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