HUMANIZED RODENTS THAT EXPRESS HEAVY CHAINS CONTAINING VL DOMAINS

Claims

1. An animal other than a human being that contains (a) insertion of one or more V gene segments L human and one or more J gene segments L human above the non-human immunoglobulin light chain constant region, (b) insertion of one or more V gene segments L human and one or more J gene segments L human above the non-human constant region of the immunoglobulin heavy chain, and (c) a nucleotide sequence that encodes an ADAM6 protein or a functional fragment thereof, wherein the ADAM6 protein is expressed from an ectopic ADAM6 nucleic acid sequence.

2. The non-human animal of claim 1, wherein the non-human heavy and / or light chain constant regions are rodent constant regions.

3. The non-human animal according to claim 1 or 2, wherein the light chain constant region is mouse Cκ.

4. A non-human animal according to any of the preceding paragraphs, in which the V gene segments L and J L The human Vκ and Jκ gene segments are located above the non-human immunoglobulin light chain constant region.

5. A non-human animal according to any of the preceding paragraphs, in which the V gene segments L and J L The human Vκ and Jκ gene segments are located above the non-human constant region of the immunoglobulin heavy chain.

6. A non-human animal according to any of the preceding claims, in which a nucleotide sequence that encodes an ADAM6 protein or a functional fragment thereof is present in the same position as a wild-type non-human ADAM6 locus.

7. A non-human animal according to any of the preceding claims, in which the nucleotide sequence that encodes the ADAM6 protein or a functional fragment thereof is present within the immunoglobulin gene segments.

8. The non-human animal of claim 7, wherein the immunoglobulin gene segments are human κ light chain gene segments.

9. The non-human animal of claim 7 or 8, wherein the immunoglobulin gene segments are endogenous heavy chain gene segments of the non-human animal.

10. A non-human animal according to any of the preceding paragraphs, wherein the non-human animal comprises endogenous V gene segments L and / or J L immunoglobulins that are incapable of rearranging to form the V domain L immunoglobulin in a non-human animal.

11. A genetically modified non-human animal that contains (a) one or more unrearranged human Vκ gene segments and one or more unrearranged human Jκ gene segments at the endogenous immunoglobulin heavy chain locus of said non-human animal, (b) one or more unrearranged human Vκ gene segments and one or more unrearranged human Jκ gene segments at an endogenous immunoglobulin light chain locus of said non-human animal, wherein said non-human animal is capable of expressing the ADAM6 protein or a functional fragment thereof.

12. A cell obtained from a non-human animal according to paragraph 1 or 11.

13. The cell of claim 12, wherein the cell is a B cell.

14. A hybridoma formed from a B cell according to claim 13.

15. The cage according to claim 12, wherein the non-human animal is a rodent.

16. The cage according to claim 15, wherein the rodent is selected from a mouse and a rat.

17. A method for producing an antigen-binding protein that binds to an antigen of interest, the method comprising (a) exposing a non-human animal to an antigen of interest according to claim 1 or 11, (b) isolating one or more B lymphocytes from a non-human animal, wherein the one or more B lymphocytes express a V binding protein L , which binds the antigen of interest, and (c) identification of the nucleic acid sequence that encodes domain V L binding protein V L , which binds the said antigen of interest, wherein the binding protein V Lcomprises a human Vκ domain and a non-human light chain constant domain and a human Vκ domain and a non-human heavy chain constant domain, and (d) using the nucleic acid sequence of (c) with a nucleic acid sequence of a human immunoglobulin constant region to produce a human antigen-binding protein that binds an antigen of interest.

18. The method of claim 17, wherein the non-human constant region of the V binding protein light chain L represents the mouse Cκ, and the non-human heavy chain constant region represents the mouse heavy chain constant region.

19. The method according to claim 17 or 18, wherein the non-human animal is a mouse.

20. A rodent comprising one or more human immunoglobulin light chain gene segments operably linked to a non-human immunoglobulin heavy chain constant region gene, wherein the rodent expresses one or more ADAM6 proteins.

21. The rodent of claim 20, wherein one or more human immunoglobulin light chain gene segments are human Vκ and Jκ gene segments.

22. The rodent of claim 20 or 21, wherein the non-human immunoglobulin heavy chain constant region gene is a mouse or rat heavy chain constant region gene.

23. The rodent of any one of claims 20-22, wherein the immunoglobulin heavy chain constant region gene comprises C H 1 and / or hinge region.

24. The rodent of any one of claims 20 to 23, wherein the rodent comprises a deletion or substitution of one or more endogenous immunoglobulin heavy chain gene sequences.

25. The rodent of any one of claims 20-24, which has endogenous V gene segments H , D H and J H are incapable of rearranging to form a rearranged V / D / J sequence.

26. The rodent of any one of claims 20-25, wherein the rodent comprises a deletion of the endogenous ADAM6 gene and further comprises an ectopic mouse ADAM6 gene.

27. A rodent according to any one of paragraphs 20-26, wherein the rodent is selected from a mouse or a rat.

28. A method for modifying the heavy chain locus of a rodent immunoglobulin, comprising: (a) making a first modification to a rodent immunoglobulin heavy chain locus that results in a reduction or elimination of endogenous ADAM6 activity in a male rodent, wherein the first modification comprises inserting a gene segment selected from one or more V gene segments L human, one or more gene segments J L humans and their combinations; and, (b) making a second modification to add a nucleic acid sequence to the rodent that provides the rodent with ADAM6 activity that is functional in a male rodent.

29. The method of claim 28, wherein the first modification further comprises inserting one or more D gene segments H humans that are capable of rearrangement with one or more V gene segments L and one or more J gene segments L .

30. The method according to claim 28, wherein one or more V gene segments L human and one or more J gene segments L human are represented by the Vκ and Jκ gene segments or the Vλ and Jλ gene segments.

31. The method of any one of claims 28-30, wherein the nucleic acid sequence that provides the rodent with ADAM6 activity that is functional in the male rodent is adjacent to one or more V gene segments L human and / or one or more J gene segments L person.

32. The method according to any one of claims 28-31, wherein the first and second modification are carried out in a separate ES cell, and the separate ES cell is introduced into the host embryo to produce a rodent.