Preparative chromatography column and methods

Abstract

A chromatography column that captures components in a process liquid in a free flow state and allows elution in steps is described.

What is claimed is: 
     
         1 . A method of chromatographic separation of a desired component comprising:
 a. Providing a disposable preparative chromatography apparatus comprising:
 i. a flexible disposable bag with an inner volume; 
 ii. a septum disposed within the flexible disposable bag to separate the flexible disposable bag into a flexible upper chamber and a flexible lower chamber, the septum including a porous surface that allows passage of liquid but not chromatography media from the upper chamber to the lower chamber; 
 iii. a chromatography media disposed in the upper chamber; 
 iv. at least one inlet to introduce liquid into the flexible upper chamber of the bag containing the chromatography media; 
 v. at least one outlet to remove the liquid from the flexible lower chamber of the bag after passage through the septum; and 
 vi. at least one mixing mechanism to mix the contents of the bag; 
   b. Introducing a quantity of liquid containing a component to be separated into the upper chamber of the flexible bag through the inlet of (iv) above;   c. Mixing the contents of the flexible bag to allow the component to bind to the chromatography media;   d. Opening the drain in the lower chamber and draining out the liquid that has passed through the septum;   e. Closing the drain;   f. Introducing into the upper chamber a solution capable of disrupting the binding between the component and the chromatography media;   g. Mixing the contents of the column until the component has been sufficiently released from the chromatography media;   h. Opening the drain in the lower chamber;   i. Collecting the contents of the flexible bag as a concentrated solution of the component.   
     
     
         2 . The method of  claim 1 , wherein the component is a protein. 
     
     
         3 . The method of  claim 1 , further comprising the use of different solutions to separate impurities from the desired component. 
     
     
         4 . The method of  claim 1 , wherein the chromatographic media after step (e) is removed from the upper chamber of the flexible bag and packed in a traditional chromatography column for further purification. 
     
     
         5 . The method of  claim 1 , wherein mixing comprises shaking, rocking or vibrating the flexible bag. 
     
     
         6 . The method of  claim 1 , wherein the chromatography media is selected from the group consisting of adsorption media, ion exchange media, hydrophobic interaction media, affinity media, immunoaffinity media and mixtures thereof.