MICE THAT MAKE VL BINDING PROTEINS
Genetically modified mice and methods for making an using them are provided, wherein the mice comprise a replacement of all or substantially all immunoglobulin heavy chain V gene segments, D gene segments, and J gene segments with at least one light chain V gene segment and at least one light chain J gene segment. Mice that make binding proteins that comprise a light chain variable domain operably linked to a heavy chain constant region are provided. 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. Modified cells, embryos, and mice that encode sequences for making the binding proteins are provided.
1 .- 21 . (canceled)
22 . A mouse whose genome comprises
a sequence that includes an unrearranged human light chain variable (V L ) gene segment and an unrearranged human light chain joining (J L ) gene segment operably linked with one another to allow for recombination to form a rearranged human V L /J L region, and further operably linked with an intact heavy chain constant (C H ) region gene,
which sequence lacks a functional D H gene segment between the unrearranged V L gene segment and the unrearranged J L gene segment, so that
when immunized with an antigen, the mouse generates an antibody in which at least one chain is comprised of an immunoglobulin light chain variable region amino acid sequence that lacks a D amino acid sequence, fused to an immunoglobulin heavy chain constant region amino acid sequence.
23 . The mouse of claim 22 , wherein the unrearranged V L and J L gene segments are Vκ and Jκ gene segments.
24 . The mouse of claim 23 , wherein the sequence replaces all of the endogenous V H , D H and J H segments at the endogenous heavy chain locus.
25 . The mouse of claim 24 , wherein:
the sequence includes a plurality of unrearranged human V L gene segments and a plurality of unrearranged human J L gene segments; and
wherein the intact C H region is in an endogenous C H region.
26 . The mouse of claim 25 , wherein the endogenous C H region is one of a plurality of endogenous C H regions.
27 . The mouse of claim 26 , wherein the sequence that includes an unrearranged human light chain variable (V L ) gene segment and an unrearranged human light chain joining (J L ) gene segment is operably linked with each of the plurality of endogenous C H regions.
28 . The mouse of claim 25 , wherein the sequence includes
(a) six human Vκ gene segments and five human Jκ gene segments;
(b) 16 human Vκ gene segments and five human Jκ gene segments;
(c) 30 human Vκ gene segments and five human Jκ gene segments; or
(d) 40 human Vκ gene segments and five human Jκ gene segments.
29 . The mouse of claim 22 , wherein the unrearranged V L and J L gene segments are Vλ and Jλ gene segments.
30 . The mouse of claim 29 , wherein the sequence replaces all of the endogenous V H , D H and J H segments at the endogenous heavy chain locus.
31 . The mouse of claim 30 , wherein
the sequence comprises a plurality of unrearranged human V L gene segments and a plurality of unrearranged human J L gene segments; and
wherein the intact C H region is an endogenous C H region.
32 . The mouse of claim 31 , wherein the sequence includes
(a) 12 human Vλ gene segments and one human Jλ gene segment;
(b) 28 human Vλ gene segments and one human Jλ gene segment; or
(c) 40 human Vλ gene segments and one human Jλ gene segment.
33 . The mouse of claim 32 , wherein the one human Jλ gene segment is human Jλ1.
34 . The mouse of claim 31 , wherein the sequence includes
(a) 12 human Vλ gene segments and four human Jλ gene segments;
(b) 28 human Vλ gene segments and four human Jλ gene segments; or
(c) 40 human Vλ gene segments and four human Jλ gene segments.
35 . The mouse of claim 34 , wherein the four human Jλ gene segments are Jλ1, Jλ2, Jλ3 and Jλ7.
36 . The mouse of claim 22 , which mouse contains B cells that comprise a rearranged V L /J L region operably linked to a mouse C H region.
37 . The mouse of claim 36 , wherein the rearranged V L /J L region is a rearranged Vκ/Jκ region.
38 . The mouse of claim 22 , which mouse, when immunized with an antigen, generates antibodies comprised of four chains:
two of which include a light chain variable region amino acid sequence fused to a light chain constant region amino acid sequence, and
two of which include an immunoglobulin light chain variable region amino acid sequence that lacks a D H amino acid sequence, fused to an immunoglobulin heavy chain constant region amino acid sequence.
39 . An isolated mouse cell whose genome comprises
a sequence that includes an unrearranged human light chain variable (V L ) gene segment and an unrearranged human light chain joining (J L ) gene segment operably linked with one another to allow for recombination to form a rearranged human V L /J L region, and further operably linked with an intact heavy chain constant (C H ) region gene,
which sequence lacks a functional D H gene segment between the unrearranged V L gene segment and the unrearranged J L gene segment.
40 . The isolated mouse cell of claim 39 , wherein the cell is an embryonic stem (ES) cell.
41 . The isolated mouse cell of claim 39 , wherein the unrearranged V L and J L gene segments are Vκ and Jκ gene segments.
42 . The isolated mouse cell of claim 39 , wherein the unrearranged V L and J L gene segments are Vλ and Jλ gene segments.
43 . The isolated mouse cell of claim 39 , wherein the intact C H region gene is a mouse C H region gene.
44 . The isolated mouse cell of claim 39 , wherein the unrearranged human V L and J L gene segments replace all of the endogenous V H , D H and J H segments at the endogenous heavy chain locus.