US2010056766A1PendingUtilityA1

Purification of biological conjugates by size exclusion chromatography

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Assignee: ABBOTT LABPriority: Aug 27, 2008Filed: Aug 27, 2008Published: Mar 4, 2010
Est. expiryAug 27, 2028(~2.1 yrs left)· nominal 20-yr term from priority
Inventors:Marcela De Leon GattiYingqing HuangDavid K. SchislaLou-Hwa J. SheuChristopher A. Tomas
C07H 17/08C07K 1/34C07G 11/00A61K 2039/505C07K 1/16
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Claims

Abstract

A method for separating a biological conjugate from an aggregate. The molecular weight of the biological conjugate ranges from about 10 kDa to about 1000 kDa. In one embodiment, the method comprises the steps of: (e) providing a mixture comprising the biological conjugate and the aggregate, wherein the biological conjugate has a molecular weight of from about 10 kDa to about 1000 kDa; (f) providing a chromatography column containing a gel, wherein the gel comprises at least one polysaccharide; (g) introducing the mixture of step (a) into the chromatography column; (h) recovering the biological conjugate from the column.

Claims

exact text as granted — not AI-modified
1 . A method for separating a biological conjugate from an aggregate comprising the steps of:
 (a) providing a mixture comprising the biological conjugate and the aggregate, wherein the biological conjugate has a molecular weight of from about 10 kDa to about 1000 kDa;   (b) providing a chromatography column containing a gel, wherein the gel comprises at least one polysaccharide;   (c) introducing the mixture of step (a) into the chromatography column;   (d) recovering the biological conjugate from the column.   
   
   
       2 . The method of  claim 1 , wherein the biological conjugate comprises a chemiluminescent label. 
   
   
       3 . The method of  claim 2 , wherein the chemiluminescent label comprises acridinium. 
   
   
       4 . The method of  claim 1 , wherein the biological conjugate comprises an antibody. 
   
   
       5 . The method of  claim 4 , wherein the antibody is an anti-biotin antibody. 
   
   
       6 . The method of  claim 1 , wherein the molecular weight of the aggregate is equal to or greater than two times the weight of a single molecule of the biological conjugate. 
   
   
       7 . The method of  claim 1 , wherein the loading volume of the chromatography column ranges from about 0.2% to about 15.4%. 
   
   
       8 . The method of  claim 1 , wherein a batch size ranges from about 50 to about 225 mg. 
   
   
       9 . The method of  claim 1 , wherein a batch size ranges from about 50 to about 150 mg. 
   
   
       10 . The method of  claim 1 , wherein the biological conjugate has a specified range of molecular structures. 
   
   
       11 . The method of  claim 10 , wherein the biological conjugate has a globular structure. 
   
   
       12 . The method of  claim 1 , wherein the input ratio ranges from about 0.5 μL of the label/mg of the specific binding member to about 6 μL of the label/mg of the specific binding member. 
   
   
       13 . The method of  claim 1 , wherein the incorporation ratio ranges from about 1.3 to about 2.5. 
   
   
       14 . The method of  claim 1 , wherein the selectivity is at least about 1.8. 
   
   
       15 . The method of  claim 15 , wherein the selectivity ranges from about 1.8 to about 3.6. 
   
   
       16 . The method of  claim 1 , wherein the at least one polysaccharide is dextran. 
   
   
       17 . The method of  claim 1 , wherein the at least one polysaccharide is agarose. 
   
   
       18 . The method of  claim 1 , wherein the at least one polysaccharide comprises dextran and agarose. 
   
   
       19 . The method of  claim 18 , wherein the dextran is covalently bound to highly cross-linked agarose.

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