Isolating cells expressing secreted proteins

Abstract

A method for identifying and isolating cells which produce secreted proteins. This method is based upon a specific characteristic or the expression level of the secreted protein by transiently capturing the secreted protein on the surface of an individual cell, allowing selection of rare cell clones from a heterogeneous population. Also provided is the use of this method to generate cells which produce a desired level of secreted protein or secreted protein of a particular characteristic(s), and organisms which possess such cells. In particular, the method allows rapid isolation of high expression recombinant antibody-producing cell lines, or may be applied directly to rapid isolation of specific hybridomas, or to the isolation of antibody-producing transgenic animals. This method is applicable for any cell which secretes protein.

1 - 86 . (canceled)  
     
     
         87 . A method of isolating an antibody, comprising 
 fusing antibody-producing cells with immortalized cells expressing a cell surface capture molecule to produce a plurality of fused cells, wherein the antibody-producing cells are B or plasma cells;    culturing the cells whereby antibody-capture molecule complexes are formed and displayed on the surface of the cells;    contacting the cells with a detection molecule that binds to an antibody an interest, whereby one or more cells bind to the detection molecule via the antibody of interest; and    isolating one or more cells that bound to the detection molecule.    
     
     
         88 . The method of  claim 87 , wherein the isolated one or more cells express antibodies with a desired specificity, isotype or avidity.  
     
     
         89 . The method of  claim 87 , wherein the capture molecule is selected from the group consisting of Fc receptors, anti-immunoglobulin antibodies, protein A, protein L, protein G and protein H, or functional fragments thereof.