Human lambda light chain mice

Abstract

Genetically modified mice are provided that express human λ variable (hVλ) sequences, including mice that express hVλ sequences from an endogenous mouse λ light chain locus, mice that express hVλ sequences from an endogenous mouse κ light chain locus, and mice that express hVλ sequences from a transgene or an episome wherein the hVλ sequence is linked to a mouse constant sequence. Mice are provided that are a source of somatically mutated human λ variable sequences useful for making antigen-binding proteins. Compositions and methods for making antigen-binding proteins that comprise human λ variable sequences, including human antibodies, are provided.

1 - 20 . (canceled) 
     
     
         21 . A method of generating a human λ light chain variable region, the method comprising:
 (a) immunizing a genetically modified mouse with an antigen, wherein the germline genome of the genetically modified mouse comprises: 
 one or more human Vλ gene segments and one or more human Jλ gene segments, wherein the one or more human Vλ gene segments and the one or more human Jλ gene segments replace endogenous mouse Vκ gene segments and endogenous Jκ gene segments; and 
 wherein the one or more human Vλ gene segments and one or more human Jλ gene segments are operably linked to an endogenous mouse κ light chain constant (Cκ) region gene; and 
 (b) determining a human λ light chain variable region that encodes a human λ light chain variable domain of an antibody that specifically binds the antigen and was generated by the genetically modified mouse, wherein the human λ light chain variable region is derived from a human Vλ gene segment of the one or more human Vλ gene segments and a human Jλ gene segment of the one or more human Jλ gene segments. 
 
     
     
         22 . The method of  claim 21 , comprising isolating a B cell of the genetically modified mouse and amplifying the λ light chain variable region from the B cell of the genetically modified mouse. 
     
     
         23 . The method of  claim 21 , wherein the one or more human Vλ gene segments and the one or more human Jλ gene segments are unrearranged gene segments. 
     
     
         24 . The method of  claim 21 , wherein the one or more human Vλ gene segments comprises at least 12 human Vλ gene segments. 
     
     
         25 . The method of  claim 21 , wherein the one or more human Vλ gene segments comprises at least 28 human Vλ gene segments. 
     
     
         26 . The method of  claim 21 , wherein the one or more human Vλ gene segments comprises at least 40 human Vλ gene segments. 
     
     
         27 . The method of  claim 21 , wherein the one or more human Jλ gene segments comprises a human Jλ1, Jλ2, Jλ3, Jλ7, or a combination thereof. 
     
     
         28 . The method of  claim 21 , wherein an endogenous mouse immunoglobulin λ light chain locus is deleted in whole or in part. 
     
     
         29 . The method of  claim 21 , wherein 10% to 45% of the B cells of the genetically modified mouse express an antibody that comprises an immunoglobulin light chain comprising a human λ light chain variable domain and a mouse Cκ domain. 
     
     
         30 . The method of  claim 29 , wherein the human λ light chain variable domain is derived from a rearranged hVλ/hJλ that is a Vλ3-1/Jλ1, Vλ3-1/Jλ7, Vλ4-3/Jλ1, Vλ4-3/Jλ7, Vλ2-8/Jλ1, Vλ3-9/Jλ1, Vλ3-10/Jλ1, Vλ3-10/Jλ3, Vλ3-10/Jλ7, Vλ2-14/Jλ1, Vλ3-19/Jλ1, Vλ2-23/Jλ1, Vλ3-25/Jλ1, Vλ1-40/Jλ1, Vλ1-40/Jλ2, Vλ1-40/Jλ3, Vλ1-40/Jλ7, Vλ7-43/Jλ1, Vλ7-43/Jλ3, Vλ1-44/Jλ1, Vλ1-44/Jλ7, Vλ5-45/Jλ1, Vλ5-45/Jλ2, Vλ5-45/Jλ7, Vλ7-48/Jλ1, Vλ7-46/Jλ2, Vλ7-48/Jλ7, Vλ9-49/Jλ1, Vλ9-49/Jλ2, Vλ9-49/Jλ7 or Vλ1/Jλ1. 
     
     
         31 . The method of  claim 21 , wherein the germline genome of the genetically modified mouse further comprises a human Vκ-Jκ intergenic region from a human immunoglobulin κ light chain locus, wherein the human Vκ-Jκ intergenic region is contiguous with a human Vλ gene segment and a human Jλ gene segment. 
     
     
         32 . The method of  claim 31 , wherein the human Vκ-Jκ intergenic region is placed between a human Vλ gene segment and a human Jλ gene segment. 
     
     
         33 . A method for generating a human λ light chain variable domain comprising:
 (a) immunizing a genetically modified mouse with an antigen, wherein the germline genome of the genetically modified mouse comprises: 
 one or more human Vλ gene segments and one or more human Jλ gene segments, wherein the one or more human Vλ gene segments and the one or more human Jλ gene segments replace endogenous mouse Vκ gene segments and endogenous Jκ gene segments; and 
 wherein the one or more human Vλ gene segments and one or more human Jλ gene segments are operably linked to an endogenous mouse Cκ gene; and 
 (b) determining a human λ light chain variable domain of an antibody that specifically binds the antigen and was generated by the genetically modified mouse. 
 
     
     
         34 . The method of  claim 33 , wherein determining a human λ light chain variable domain comprises determining a nucleotide sequence that encodes the human λ light chain variable domain. 
     
     
         35 . The method of  claim 33 , wherein the one or more human Vλ gene segments and one or more human Jλ gene segments are unrearranged gene segments. 
     
     
         36 . The method of  claim 33 , wherein the one or more human Vλ gene segments comprises at least 12 human Vλ gene segments. 
     
     
         37 . The method of  claim 33 , wherein the one or more human Vλ gene segments comprises at least 28 human Vλ gene segments. 
     
     
         38 . The method of  claim 33 , wherein the one or more human Vλ gene segments comprises at least 40 human Vλ gene segments. 
     
     
         39 . The method of  claim 33 , wherein the one or more human Jλ gene segments comprises a human Jλ1, Jλ2, Jλ3, Jλ7, or a combination thereof. 
     
     
         40 . The method of  claim 33 , wherein an endogenous mouse immunoglobulin λ light chain locus is deleted in whole or in part. 
     
     
         41 . The method of  claim 33 , wherein 10% to 45% of the B cells of the genetically modified mouse express an antibody that comprises an immunoglobulin light chain comprising a human λ light chain variable domain and a mouse Cκ domain. 
     
     
         42 . The method of  claim 41 , wherein the human λ light chain variable domain is derived from a rearranged hVλ/hJλ that is a Vλ3-1/Jλ1, Vλ3-1/Jλ7, Vλ4-3/Jλ1, Vλ4-3/Jλ7, Vλ2-8/Jλ1, Vλ3-9/Jλ1, Vλ3-10/Jλ1, Vλ3-10/Jλ3, Vλ3-10/Jλ7, Vλ2-14/Jλ1, Vλ3-19/Jλ1, Vλ2-23/Jλ1, Vλ3-25/Jλ1, Vλ1-40/Jλ1, Vλ1-40/Jλ2, Vλ1-40/Jλ3, Vλ1-40/Jλ7, Vλ7-43/Jλ1, Vλ7-43/Jλ3, Vλ1-44/Jλ1, Vλ1-44/Jλ7, Vλ5-45/Jλ1, Vλ5-45/Jλ2, Vλ5-45/Jλ7, Vλ7-48/Jλ1, Vλ7-46/Jλ2, Vλ7-48/Jλ7, Vλ9-49/Jλ1, Vλ9-49/Jλ2, Vλ9-49/Jλ7 or Vλ1-51/Jλ1. 
     
     
         43 . The method of  claim 33 , wherein the germline genome of the genetically modified mouse further comprises a human Vκ-Jκ intergenic region from a human immunoglobulin κ light chain locus, wherein the human Vκ-Jκ intergenic region is contiguous with a human V gene segment and a human Jλ gene segment. 
     
     
         44 . The method of  claim 43 , wherein the human Vκ-Jκ intergenic region is placed between a human Vλ gene segment and a human Jλ gene segment.