Methods for obtaining antibodies

Abstract

The invention is directed towards a method of enriching a population of cells in those cells that produce an antibody which recognises an antigen of interest. In particular, an untagged antigen is used in conjunction with a polyclonal antibody to isolate cells recognizing said antigen.

1 . A method of enriching a population of cells in those cells which produce an antibody that recognises an antigen of interest, comprising: 
 a) bringing said population into contact with an antibody that recognises a marker which is essentially unique to those cells present in the population which are capable of producing an antibody, said antibody being attached to a first fluorescent label;    b) bringing said population into contact with the antigen of interest;    c) bringing said population into contact with a sample comprising an antibody that recognises said antigen, said antibody being attached to a second fluorescent label; and    d) separating from the population those cells which are detectable by virtue of being associated with the first and second fluorescent labels.    
     
     
         2 . The method of  claim 1 , wherein parts a), b), and c) are performed simultaneously and the performance optionally comprises at least one wash step.  
     
     
         3 . The method of  claim 1 , wherein parts a) and b), or a) and c), or b) and c), are performed simultaneously and optionally comprise at least one wash step.  
     
     
         4 . The method of  claim 1 , wherein parts a), b) and c) are performed consecutively in any order, and wherein each performance optionally comprises at least one wash step.  
     
     
         5 . The method of  claim 1 , wherein part a) additionally comprises bringing said population into contact with an antibody that recognises a second marker essentially unique to those cells present in the population which are capable of producing an antibody, said antibody being labelled with a third label.  
     
     
         6 . The method of  claim 1 , wherein the separation of the cells producing an antibody that recognises the antigen of interest is performed using fluorescence activated cell sorting.  
     
     
         7 . A method according to  claim 1  additionally comprising: 
 a) culturing a plurality of those cells associated with the antigen-antibody-particle complex;    b) screening the cultured cells to identify those cells capable of producing an antibody which recognises an antigen of interest; and    c) isolating said antibody directly or indirectly from the cells.    
     
     
         8 . An antibody prepared directly or indirectly according to the method of  claim 1 .  
     
     
         9 . A pharmaceutical composition comprising an antibody according to  claim 8  and a pharmaceutically acceptable carrier.  
     
     
         10 . (canceled)  
     
     
         11 . (canceled)  
     
     
         12 . The method of  claim 2 , wherein the separation of the cells producing an antibody that recognises the antigen of interest is performed using fluorescence activated cell sorting.  
     
     
         13 . The method of  claim 4 , wherein the separation of the cells producing an antibody that recognises the antigen of interest is performed using fluorescence activated cell sorting.  
     
     
         14 . The method of  claim 5 , wherein the separation of the cells producing an antibody that recognises the antigen of interest is performed using fluorescence activated cell sorting.  
     
     
         15 . A method according to  claim 2  additionally comprising: 
 a) culturing a plurality of those cells associated with the antigen-antibody-particle complex;    b) screening the cultured cells to identify those cells capable of producing an antibody which recognises an antigen of interest; and    c) isolating said antibody directly or indirectly from the cells.    
     
     
         16 . A method according to  claim 4  additionally comprising: 
 a) culturing a plurality of those cells associated with the antigen-antibody-particle complex;    b) screening the cultured cells to identify those cells capable of producing an antibody which recognises an antigen of interest; and    c) isolating said antibody directly or indirectly from the cells.    
     
     
         17 . A method according to  claim 5  additionally comprising: 
 a) culturing a plurality of those cells associated with the antigen-antibody-particle complex;    b) screening the cultured cells to identify those cells capable of producing an antibody which recognises an antigen of interest; and    c) isolating said antibody directly or indirectly from the cells.    
     
     
         18 . A method according to  claim 6  additionally comprising: 
 a) culturing a plurality of those cells associated with the antigen-antibody-particle complex;    b) screening the cultured cells to identify those cells capable of producing an antibody which recognises an antigen of interest; and    c) isolating said antibody directly or indirectly from the cells.    
     
     
         19 . An antibody prepared directly or indirectly according to the method of  claim 5 .  
     
     
         20 . An antibody prepared directly or indirectly according to the method of  claim 7.