Recombinant production of mixtures of antibodies

Abstract

Provided is methods for producing mixtures of antibodies from a single host cell clone, wherein, a nucleic acid sequence encoding a light chain and nucleic acid sequences encoding different heavy chains are expressed in a recombinant host cell. The recombinantly produced antibodies in the mixtures according to the invention suitably comprise identical light chains paired to different heavy chains capable of pairing to the light chain, thereby forming functional antigen-binding domains. Mixtures of the recombinantly produced antibodies are also provided by the invention. Such mixtures can be used in a variety of fields.

1 .- 92 . (canceled) 
     
     
         93 . An isolated host cell comprising an exogenously introduced nucleic acid sequence encoding an immunoglobulin light chain variable region and three or more immunoglobulin heavy chain variable regions. 
     
     
         94 . The isolated host cell of  claim 93 , wherein the three or more immunoglobulin heavy chain variable regions are capable of pairing with the immunoglobulin light chain variable region to form a multispecific immunoglobulin. 
     
     
         95 . The isolated host cell of  claim 94 , wherein the multispecific immunoglobulin comprises an antigen-binding antibody fragment. 
     
     
         96 . The isolated host cell of  claim 95 , wherein the antibody fragment is a Fv, Fab, Fab′, or F(ab′) 2  fragment. 
     
     
         97 . The isolated host cell of  claim 93 , which expresses an immunoglobulin comprising two antigen-binding domains. 
     
     
         98 . The isolated host cell of  claim 97 , wherein said antigen-binding domains are not identical. 
     
     
         99 . The isolated host cell of  claim 93 , wherein the three or more immunoglobulin heavy chain variable regions are capable of paring with immunoglobulin light chain variable region to form three or more non-identical antigen-binding antibody fragments. 
     
     
         100 . The isolated host cell of  claim 93 , wherein the immunoglobulin heavy chain variable regions have different specificities and/or affinities. 
     
     
         101 . The isolated host cell of  claim 93 , wherein the immunoglobulin heavy chain variable regions bind to different epitopes on the same antigen. 
     
     
         102 . The isolated host cell of  claim 93 , wherein the immunoglobulin heavy chain variable regions bind to different antigens. 
     
     
         103 . The isolated host cell of  claim 93 , wherein the exogenously introduced nucleic acid sequence comprises a tissue-specific promoter. 
     
     
         104 . The isolated host cell of  claim 93 , wherein the exogenously introduced nucleic acid sequence is integrated into the host cell's genome. 
     
     
         105 . The isolated host cell of  claim 104 , wherein the exogenously introduced nucleic acid sequence is stably integrated at a predetermined position. 
     
     
         106 . The isolated host cell of  claim 105 , wherein the integration is performed using homologous recombination or site-specific recombinases. 
     
     
         107 . The isolated host cell of  claim 93 , wherein the nucleic acid sequence encoding an immunoglobulin light chain variable region and the three or more immunoglobulin heavy chain variable regions are introduced into the host cell consecutively. 
     
     
         108 . The isolated host cell of  claim 93 , wherein the nucleic acid sequence encoding an immunoglobulin light chain variable region and the three or more immunoglobulin heavy chain variable regions are introduced into the host cell concomitantly. 
     
     
         109 . A cell culture comprising the host cell of  claim 93  and a culture medium. 
     
     
         110 . The cell culture of  claim 109 , which produces two or more non-identical antibodies for more than 20 population doublings. 
     
     
         111 . The isolated host cell of  claim 93 , wherein said encoded immunoglobulin light chain variable region and three or more immunoglobulin heavy chain variable regions are encoded by a single nucleic acid. 
     
     
         112 . The isolated host cell of  claim 93 , wherein said encoded immunoglobulin light chain variable region and three or more immunoglobulin heavy chain variable regions are encoded by one or more nucleic acids.