Method of cloning a lambda light chain variable domain using a human lambda light chain mouse
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. A method for cloning a λ light chain variable domain, comprising the steps of:
(a) obtaining a cell from a transgenic mouse whose genome comprises an endogenous κ light chain immunoglobulin locus comprising a replacement of endogenous Vκ and Jκ gene segments with human Vλ and Jλ gene segments, wherein the human Vλ and Jλ gene segments are operably linked to a mouse Cκ gene such that the mouse expresses an immunoglobulin light chain that comprises a human λ variable region nucleic acid sequence fused with a mouse κ constant region nucleic acid sequence;
(b) cloning a λ light chain variable domain derived from the human Vλ and Jλ gene segments.
2. The method of claim 1 , wherein the cell is a hybridoma.
3. The method of claim 1 , wherein the cell is a quadroma.
4. The method of claim 1 , wherein the cell is a B cell.
5. The method of claim 1 , wherein the cell has a genome that comprises one or more human heavy chain V, D, and J segments.
6. The method of claim 5 , wherein the one or more human heavy chain V, D, and J segments are at an endogenous mouse heavy chain locus.
7. The method of claim 1 , wherein the cell has a genome comprising a replacement at the endogenous mouse heavy chain variable locus of one or more endogenous heavy chain V, D, and J segments with one or more human heavy chain V, D, and J segments.
8. The method of claim 5 , further comprising cloning from the cell a human heavy chain variable (V H ) region nucleic acid sequence that is cognate with the human λ variable region nucleic acid sequence.
9. The method of claim 8 , further comprising employing the nucleic acid sequences of the human V H and human λ variable regions fused to human constant regions to make a human antibody.
10. The method of claim 1 , wherein the human Vλ and Jλ gene segments comprise at least 12 human Vλ gene segments.
11. The method of claim 1 , wherein the human Vλ and Jλ gene segments comprise at least 28 hVλ gene segments.
12. The method of claim 1 , wherein human Vλ and Jλ gene segments comprise at least 40 human Vλ gene segments.
13. The method of claim 1 , wherein the human Vλ and Jλ gene segments comprise a human Jλ1 gene segment.
14. The method of claim 1 , wherein the mouse further comprises a human Vκ-Jκ intergenic region sequence located between the human Vλ gene segments and the human Jλ gene segments, wherein the Vκ-Jκ intergenic region is the region located about 130 bp downstream of the 3′ untranslated region of a human Vκ4-1 gene segment to about 600 bp upstream of a human Jκ1 gene segment as found in a human genome.
15. The method of claim 14 , wherein the human Vκ-Jκ intergenic region sequence comprises SEQ ID NO: 100.