Vl antigen binding proteins exhibiting distinct binding characteristics

Abstract

Methods for making, identifying, isolating and/or making binding proteins that contain an immunoglobulin light chain variable domain, including a somatically hypermutated light chain variable domain, fused with a heavy chain constant region, are provided. Exemplary binding proteins specific to small molecules are also provided.

1 .- 20 . (canceled) 
     
     
         21 . A genetically modified non-human animal comprising
 (a) in its genome
 (i) unrearranged human immunoglobulin light chain variable (V L ) and light chain joining (J L ) gene segments operably linked to a non-human heavy chain constant region nucleic acid sequence, and 
 (ii) unrearranged human immunoglobulin light chain variable (V L ) and light chain joining (J L ) gene segments operably linked to a non-human light chain constant region nucleic acid sequence; and 
   (b) a V L  antigen binding protein that specifically binds a small molecule.   
     
     
         22 . The genetically modified non-human animal of  claim 21 , wherein the non-human animal exhibits a 2-fold or more increase in small molecule specific binding protein titer as compared to a reference non-human animal. 
     
     
         23 .- 31 . (canceled) 
     
     
         32 . The genetically modified non-human animal of  claim 21 , wherein the non-human animal is a mouse, and the mouse comprises
 (a) in its genome
 (i) a first nucleotide sequence comprising
 (A) at least an unrearranged human immunoglobulin light chain variable kappa (Vκ) 4-1 gene segment, an unrearranged human Vκ1-5 gene segment, and/or an unrearranged human Vκ3-15 gene segment, and 
 (B) all five unrearranged human light chain joining kappa (Jκ1, Jκ2, Jκ3, Jκ4, and Jκ5) gene segments, and 
 operably linked to a mouse immunoglobulin heavy chain constant region nucleic acid sequence, and 
 
 (ii) a second nucleotide sequence comprising
 (A) at least an unrearranged human immunoglobulin light chain Vκ3-20 gene segment and/or an unrearranged human Vκ1-39 gene segment, and 
 (B) all five unrearranged human Jκ1, Jκ2, Jκ3, Jκ4, and Jκ5 gene segments, and 
 operably linked to a mouse immunoglobulin light chain constant region nucleic acid sequence, 
 
   (b) a Vκ antigen-binding protein, or a cell expressing the Vκ antigen-binding protein, wherein the Vκ antigen-binding protein comprises a first and a second immunoglobulin light chain Vκ variable domain, wherein the first immunoglobulin light chain Vκ variable domain is operably linked to an immunoglobulin heavy chain constant domain, wherein the second immunoglobulin light chain κ variable domain is operably linked to an immunoglobulin light chain κ constant domain, and wherein the first and the second immunoglobulin light chain κ variable domains have heterogeneous sequences and are associated to form a binding pocket of the Vκ antigen-binding protein that specifically binds a steroid with a K D  of about 4.4 nM to 1.93 μM, and   (c) the steroid or the steroid linked to a carrier.   
     
     
         33 . The non-human animal of  claim 32 , wherein the steroid is a hormone, an alkaloid, or cardiotonic steroid. 
     
     
         34 . The non-human animal of  claim 32 , wherein the steroid linked to a carrier is a hapten. 
     
     
         35 . The non-human animal of  claim 33 , wherein the steroid linked to a carrier is a hapten. 
     
     
         36 . The non-human animal of  claim 32 , wherein the binding pocket of the Vκ antigen-binding protein specifically binds the steroid with a K D  of 10 nM or less. 
     
     
         37 . The non-human animal of  claim 33 , wherein the binding pocket of the Vκ antigen-binding protein specifically binds the steroid with a K D  of 10 nM or less. 
     
     
         38 . The non-human animal of  claim 34 , wherein the binding pocket of the Vκ antigen-binding protein specifically binds the steroid with a K D  of 10 nM or less. 
     
     
         39 . The non-human animal of  claim 35 , wherein the binding pocket of the Vκ antigen-binding protein specifically binds the steroid with a K D  of 10 nM or less. 
     
     
         40 . The non-human animal of  claim 32 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         41 . The non-human animal of  claim 33 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         42 . The non-human animal of  claim 34 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         43 . The non-human animal of  claim 35 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         44 . The non-human animal of  claim 36 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         45 . The non-human animal of  claim 37 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         46 . The non-human animal of  claim 45 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment. 
     
     
         47 . The non-human animal of  claim 46 , wherein the first nucleotide sequence comprises an unrearranged human Vκ 4-1 gene segment, an unrearranged human Vκ 5-2 gene segment, an unrearranged human Vκ 7-3 gene segment, an unrearranged human Vκ 2-4 gene segment, an unrearranged human Vκ 1-5 gene segment, an unrearranged human Vκ 1-6 gene segment, an unrearranged human Vκ 3-7 gene segment, an unrearranged human Vκ 1-8 gene segment, an unrearranged human Vκ 1-9 gene segment, an unrearranged human Vκ 2-10 gene segment, an unrearranged human Vκ 3-11 gene segment, an unrearranged human Vκ 1-12 gene segment, an unrearranged human Vκ 1-13 gene segment, an unrearranged human Vκ 2-14 gene segment, an unrearranged human Vκ 3-15 gene segment, an unrearranged human Vκ 1-16 gene segment, an unrearranged human Vκ 1-17 gene segment, an unrearranged human Vκ 2-18 gene segment, an unrearranged human Vκ 2-19 gene segment, an unrearranged human Vκ 3-20 gene segment, an unrearranged human Vκ 6-21 gene segment, an unrearranged human Vκ 1-22 gene segment, an unrearranged human Vκ 1-23 gene segment, an unrearranged human Vκ 2-24 gene segment, an unrearranged human Vκ 3-25 gene segment, an unrearranged human Vκ 2-26 gene segment, an unrearranged human Vκ 1-27 gene segment, an unrearranged human Vκ 2-28 gene segment, an unrearranged human Vκ 2-29 gene segment, an unrearranged human Vκ 2-30 gene segment, an unrearranged human Vκ 3-31 gene segment, an unrearranged human Vκ 1-32, gene segment an unrearranged human Vκ 1-33 gene segment, an unrearranged human Vκ 3-34 gene segment, an unrearranged human Vκ 1-35 gene segment, an unrearranged human Vκ 2-36 gene segment, an unrearranged human Vκ 1-37 gene segment, an unrearranged human Vκ 2-38 gene segment, an unrearranged human Vκ 1-39 gene segment, and an unrearranged human Vκ 2-40 gene segment.