Whole cell selection utilizing azlactone-functional supports

Abstract

A method for cell selection is disclosed. The method uses an azlactone-functional support, which is derivatized with and covalently coupled to a substance that is biologically active with a desired type of whole cell. The method allows the whole cells in the mixture to interact with and bind to the coupled biologically active substance, after which a remainder of the mixture is removed from the support. Optionally, one can elute the bound whole cells from the coupled biologically active substance to produce a purified collection of the whole cells.

What is claimed is:  
     
         1 . A method for cell selection comprising the steps of: 
 (a) providing an aziactone-functional support,    (b) derivatizing the azlactone-functional support with a substance that is biologically active with a desired type of whole cell, wherein the substance is covalently coupled to the azlactone-functional support,    (c) contacting the product of step (b) with a mixture containing the whole cells,    (d) allowing the whole cells in the mixture to interact with and bind to the coupled biologically active substance,    (e) removing a remainder of the mixture from the support, and    (i) optionally, eluting the bound cells from the coupled biologically active substance to produce a purified collection of the whole cells.    
     
     
         2 . The method of  Claim 1 , wherein the azlactone-functional support is selected from the group consisting of a bead, a particulate, a membrane, a blended article, a graft copolymeric article, a woven web, a nonwoven web, a solid plastic article having a surface comprising azlactone moieties, and combinations thereof.  
     
     
         3 . The method of  claim 1 , wherein the biologically active substance is selected from the group consisting of antibodies, lectins, proteins, antigens, avidin, and combinations thereof.  
     
     
         4 . The method of  claim 1 , wherein the biologically active substance directly interacts with the whole cells.  
     
     
         5 . The method of  claim 1 , wherein the biologically active substance indirectly interacts with the whole cells through a second, intermediary biologically active substance that is bifunctional to both the whole cells and the azlactone-functional support.  
     
     
         6 . The method of  claim 1 , wherein the azlactone-functional support is prepared by processes selected from the group consisting of suspension polymerization processes and dispersion polymerization processes.  
     
     
         7 . The method of  claim 6 , wherein the aziactone-functional support is prepared from 2-alkenyl azlactone monomers and, optionally, comonomers and crosslinkers.  
     
     
         8 . The method of  claim 2 , wherein the solid plastic article is a microtitration well, a microtitration plate, a petri dish, medical tubing, a test tube, a centrifuge tube, a beaker, a cuvette, or a body implant.  
     
     
         9 . The method of  claim 1 , wherein the optional step (f) is used for further biological processing of the whole cells.  
     
     
         10 . The method of  claim 1 , wherein the mixture is selected from the group consisting of bone marrow and peripheral blood.  
     
     
         11 . A purified whole cell population produced by the method of  claim 1 .  
     
     
         12 . An interacted support, comprising: 
 (a) an azlactone-functional support,    (b) a biologically active substance covalently coupled to the support, and    (c) a whole cell interacting with said substance.    
     
     
         13 . The support of  claim 12 , wherein the wherein the azlactone-functional support is selected from the group consisting of a bead, a particulate, a membrane, a blended article, a graft copolymeric article, a woven web, a nonwoven web, a solid plastic article having a surface comprising azlactone moieties, and combinations thereof.  
     
     
         14 . The support of  claim 12 , wherein the biologically active substance is selected from the group consisting of antibodies, lectins, proteins, antigens, avidin, and combinations thereof.  
     
     
         15 . The support of  claim 12 , wherein the biologically active substance indirectly interacts with the whole cells through a second, intermediary biologically active substance that is bifunctional to both the whole cells and the azlactone-functional support.  
     
     
         16 . The support of  claim 13 , wherein the solid plastic article is a microtitration well, a microtitration plate, a petri dish, medical tubing, a test tube, a centrifuge tube, a beaker, a cuvette, or a body implant.  
     
     
         17 . The support of  claim 12 , wherein the aziactone-functional support prior to covalent coupling with the biologically active substance has at least one azlactone-functional group of a formula:  
       
         
           
           
               
               
           
         
       
       wherein: 
 R 1  and R 2  independently can be an alkyl group having 1 to 14 carbon atoms, a cycloalkyl group having 3 to 14 carbon atoms, an aryl group having 5 to 12 ring atoms, an arenyl group having 6 to 26 carbon atoms and 0 to 3 S, N, and nonperoxidic 0 heteroatoms, or R 1  and R 2  taken together with the carbon to which they are joined can form a carbocyclic ring containing 4 to 12 ring atoms, and n is an integer 0 or 1.