Antibody producing non-human animals

Abstract

Described are transgenic, non-human animals comprising a nucleic acid encoding an immunoglobulin light chain, whereby the immunoglobulin light chain is a common human, human-like, or humanized light chain. Further provided is methods for producing an immunoglobulin from the transgenic, non-human animal.

1 - 19 . (canceled) 
     
     
         20 . A method for selecting a single recombinant cell that expresses a combination of antibodies, or antibody fragments thereof, wherein the antibodies, or antibody fragments thereof, are human, humanized, or deimmunized, and wherein the combination of antibodies has specific affinity for at least two target epitopes, the method comprising:
 providing at least three different variable regions comprising two heavy chain variable regions and one light chain variable region in recombinant cells, wherein the at least three different variable regions are encoded by nucleic acid sequences identified from a transgenic non-human animal, and the antigen binding parts of the variable regions originate from a single species and wherein one variable region is able to functionally pair with more than one other variable region,   pairing of variable regions and secretion of the paired regions from the recombinant cells resulting in the production of said combination of antibodies,   providing at least two target epitopes; and   selecting a single recombinant cell from the recombinant cells that produces a combination of antibodies that binds the two target epitopes,   
       wherein the one light chain variable region comprises a rearranged human immunoglobulin light chain variable region encoded by a human light chain V gene segment and J gene segment. 
     
     
         21 . The method according to  claim 20 , wherein said at least two target epitopes are associated with a disease and/or disorder. 
     
     
         22 . The method according to  claim 21 , further comprising subjecting the combination of antibodies to a biological assay indicative of an effect of the combination of antibodies on the disease and/or disorder. 
     
     
         23 . The method according to  claim 20 , wherein the one variable region able to functionally pair with more than one other variable region does not significantly contribute to the resulting binding specificity of the resulting paired regions. 
     
     
         24 . The method according to  claim 20 , wherein each variable region can only pair with one other variable region. 
     
     
         25 . The method according to  claim 20 , wherein two of the at least three variable regions are part of one single chain Fv. 
     
     
         26 . The method according to  claim 20 , wherein the expression of two of the variable regions is under the direction of different control elements. 
     
     
         27 . The method according to  claim 26 , wherein the different control elements lead to differential expression. 
     
     
         28 . The method according to  claim 27 , wherein the differential expression is different in levels of expression and/or time of expression. 
     
     
         29 . The method according to  claim 20 , wherein the combination of antibodies comprises two monospecific antibodies produced in the single recombinant cell. 
     
     
         30 . The method according to  claim 20 , wherein the combination of antibodies comprises monospecific and bispecific antibodies produced in the single recombinant cell. 
     
     
         31 . The method according to  claim 20 , further comprising:
 producing an expression system comprising polynucleotides encoding variable regions, said producing comprising:
 synthesizing polynucleotides encoding variable regions, 
 expressing said polynucleotides and allowing the expression products to pair; and 
 selecting polynucleotides encoding variable regions having desired pairing behavior, so as to produce polynucleotides encoding variable regions. 
   
     
     
         32 . The method according to  claim 20 , further comprising:
 expressing the combination of antibodies, or antibody fragments thereof, resulting in the production of said combination of antibodies in the single recombinant cell.   
     
     
         33 . The method according to  claim 20 , wherein the human immunoglobulin light chain V gene segment and J gene segment is identified from a transgenic non-human animal. 
     
     
         34 . The method according to  claim 33 , wherein the at least three different variable regions comprising two heavy chain regions and one light chain variable region are encoded by nucleic acid sequences identified from a transgenic non-human animal. 
     
     
         35 . The method according to  claim 34 , wherein the genome of the transgenic non-human animal comprises a transgene comprising the human immunoglobulin light chain V gene segment and J segment,
 wherein the human V/J gene segments encode the rearranged human immunoglobulin light chain variable region; and   wherein the transgenic non-human animal, in response to the at least two target epitopes, produces antibodies with immunoglobulin light chains comprising the rearranged human light chain variable region and an animal light chain constant region, paired with a diversity of immunoglobulin heavy chains which bind the at least two target epitopes.   
     
     
         36 . The method according to  claim 20 , wherein a first heavy chain of the two heavy chain regions is identified by immunizing a first non-human animal with a first target epitope of the at least two target epitopes and wherein a second heavy chain of the two heavy chain regions is identified by immunizing a second non-human animal with a second target epitope of the at least two target epitopes. 
     
     
         37 . The method according to  claim 20 ,
 wherein the genome of the transgenic non-human animal comprises a transgene comprising the human immunoglobulin light chain V gene segment and J gene segment,   wherein the human V/J gene segments encode the rearranged human immunoglobulin light chain variable region; and   wherein the transgenic non-human animal, in response to the at least two target epitopes, produces antibodies with immunoglobulin light chains comprising the rearranged human light chain variable region and an animal light chain constant region, paired with a diversity of immunoglobulin heavy chains which bind the at least two target epitopes.