Histidine engineered light chain antibodies and genetically modified non-human animals for generating the same

1. A genetically modified non-human animal comprising in its germline an immunoglobulin light chain locus that comprises a single rearranged human immunoglobulin light chain variable region gene sequence comprising human Vκ and Jκ segment sequences,

wherein the Vκ segment sequence is derived from a human Vκ1-39 or Vκ3-20 gene segment, and

wherein the single rearranged human immunoglobulin light chain variable region gene sequence comprises a substitution of at least one non-histidine codon of the Vκ segment sequence with a histidine codon that is expressed at a position selected from the group consisting of 105, 106, 107, 108, 109, 111 and a combination thereof (according to IMGT numbering).

2. The animal of claim 1 , wherein the single rearranged human immunoglobulin variable region sequence is operably linked to an immunoglobulin light chain constant region gene sequence.

3. The animal of claim 2 , wherein the immunoglobulin light chain constant region gene sequence is a non-human immunoglobulin light chain constant region gene sequence.

4. The animal of claim 3 , wherein the non-human immunoglobulin light chain constant region gene sequence is an endogenous non-human immunoglobulin light chain constant region gene sequence.

5. The animal of claim 1 , further comprising in its germline an immunoglobulin heavy chain locus that comprises an unrearranged immunoglobulin heavy chain variable region gene sequence comprising human V H , D H , and J H segments operably linked to an immunoglobulin heavy chain constant region gene sequence.

6. The animal of claim 5 , wherein the immunoglobulin heavy chain constant region gene sequence is a non-human immunoglobulin heavy chain constant region gene sequence.

7. The animal of claim 6 , wherein the non-human heavy chain constant region gene sequence is an endogenous non-human immunoglobulin constant region gene sequence.

8. The animal of claim 1 , wherein the animal lacks a functional unrearranged immunoglobulin κ light chain variable region.

9. The animal of claim 1 , wherein the immunoglobulin light chain locus is at an endogenous non-human immunoglobulin light chain locus.

10. The animal of claim 5 , wherein the immunoglobulin heavy chain locus is at an endogenous non-human immunoglobulin heavy chain locus.

11. The animal of claim 1 , wherein the substitution of at least one non-histidine codon with a histidine codon is in the nucleotide sequence encoding a complementary determining region (CDR) 3.

12. The animal of claim 11 , wherein the substitution is of one, two, three, or four CDR3 codon(s).

13. The animal of claim 1 , wherein the single rearranged human immunoglobulin light chain variable region is derived from a rearranged Vκ1-39/Jκ5 or Vκ3-20/Jκ1 gene sequence.

14. The animal of claim 13 , wherein the single rearranged human immunoglobulin light chain variable region is derived from a rearranged Vκ1-39/Jκ5 gene sequence, and wherein the Vκ1-39/Jκ5 gene sequence comprises a replacement of at least one non-histidine codon with a histidine codon designed to express a histidine at a position selected from the group consisting of 105, 106, 108, 111, and a combination thereof (according to IMGT numbering).

15. The animal of claim 13 , wherein the single rearranged human immunoglobulin light chain variable region is derived from a rearranged Vκ3-20/Jκ1 gene sequence, and wherein the Vκ3-20/Jκ1 gene sequence comprises a replacement of at least one non-histidine codon with a histidine codon designed to express a histidine at position selected from the group consisting of 105, 106, 107, 109, and a combination thereof (according to IMGT numbering).

16. The animal of claim 1 , wherein the animal comprises a population of B cells in response to an antigen of interest that is enriched for antibodies that exhibit a decrease in dissociative half-life (t 1/2 ) at an acidic pH as compared to neutral pH of at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 10-fold, at least about 15-fold, at least about 20-fold, at least about 25-fold, or at least about 30-fold.

17. The animal of claim 16 , wherein the decrease in dissociative half-life (t 1/2 ) at an acidic pH as compared to neutral pH is about 30 fold or more.

18. The animal of claim 1 , wherein the animal is a rodent.

19. The rodent of claim 18 , wherein the rodent is a rat or a mouse.

20. The rodent of claim 19 , wherein the rodent is a mouse.

21. The animal of claim 1 , wherein the animal expresses an antibody comprising a human immunoglobulin light chain variable domain with a substitution of at least one non-histidine residue with a histidine at an amino acid position encoded by the at least one codon substituted in the immunoglobulin light chain variable region gene sequence.

22. A genetically modified mouse comprising in its germline an immunoglobulin light chain locus that comprises a single rearranged human immunoglobulin light chain variable region gene sequence comprising human Vκ and Jκ segment sequences,

wherein the Vκ segment sequence is derived from a human Vκ1-39 or Vκ3-20 gene segment, and

wherein the single rearranged human immunoglobulin light chain variable region sequence comprises a substitution of at least one non-histidine codon of the Vκ segment sequence with a histidine codon that is expressed at a position selected from the group consisting of 105, 106, 107, 108, 109, 111 and a combination thereof (according to IMGT numbering), and

wherein the mouse lacks a functional unrearranged immunoglobulin κ light chain variable region.

23. The mouse of claim 22 , wherein the single rearranged immunoglobulin light chain variable region gene sequence is operably linked to an immunoglobulin light chain constant region gene sequence.

24. The mouse of claim 23 , wherein the immunoglobulin light chain constant region gene sequence is selected from a rat or a mouse immunoglobulin light chain constant region gene sequence.

25. The mouse of claim 22 , further comprising in its germline an immunoglobulin heavy chain locus that comprises an unrearranged immunoglobulin heavy chain variable region sequence comprising human V H , D H , and J H segments operably linked to an immunoglobulin heavy chain constant region gene sequence.

26. The mouse of claim 25 , wherein the immunoglobulin heavy chain constant region gene sequence is a rat or a mouse heavy chain constant region gene sequence.

27. The mouse of claim 22 , wherein the immunoglobulin light chain locus is at an endogenous mouse immunoglobulin light chain locus.

28. The mouse of claim 25 , wherein the immunoglobulin heavy chain locus is at an endogenous mouse immunoglobulin heavy chain locus.

29. The mouse of claim 22 , wherein the substitution of at least one non-histidine codon with a histidine codon is in the nucleotide sequence encoding a complementary determining region (CDR) 3.

30. The mouse of claim 29 , wherein the substitution is of one, two, three, or four CDR3 codon(s).

31. The mouse of claim 22 , wherein the single rearranged human immunoglobulin light chain variable region is derived from a rearranged Vκ1-39/Jκ5 or Vκ3-20/Jκ1 gene sequence.

32. The mouse of claim 31 , wherein the single rearranged human immunoglobulin light chain variable region is derived from a rearranged Vκ1-39/Jκ5 gene sequence, and wherein the Vκ1-39/Jκ5 gene sequence comprises a replacement of at least one non-histidine codon with a histidine codon designed to express a histidine at a position selected from the group consisting of 105, 106, 108, 111, and a combination thereof (according to IMGT numbering).

33. The mouse of claim 32 , wherein the Vκ1-39/Jκ5 gene sequence comprises a replacement of non-histidine codons designed to express histidines at positions 105, 106, 108, and 111 (according to IMGT numbering).

34. The mouse of claim 32 , wherein the Vκ1-39/Jκ5 gene sequence comprises a replacement of non-histidine codons designed to express histidines at positions 106, 108 and 111 (according to IMGT numbering).

35. The mouse of claim 31 , wherein the single rearranged human immunoglobulin light chain variable region is derived from a rearranged Vκ3-20/Jκ1 gene sequence, and wherein the Vκ3-20/Jκ1 gene sequence comprises replacement of at least one non-histidine codon with a histidine codon designed to express a histidine at a position selected from the group consisting of 105, 106, 107, 109, and a combination thereof (according to IMGT numbering).

36. The mouse of claim 35 , wherein the Vκ3-20/Jκ1 gene sequence comprises a replacement of non-histidine codons designed to express histidines at positions 105, 106, 107, and 109 (according to IMGT numbering).

37. The mouse of claim 35 , wherein the Vκ3-20/Jκ1 gene sequence comprises a replacement of non-histidine codons designed to express histidines at IGMT positions 105, 106 and 109 (according to IMGT numbering).

38. The mouse of claim 22 , wherein the mouse expresses a population of antigen-specific antibodies in response to an antigen of interest wherein all antibodies comprise:

immunoglobulin light chain variable domains derived from the same single rearranged human light chain variable region gene sequence which comprises a substitution of at least one non-histidine codon with a histidine codon, and

immunoglobulin heavy chains comprising human heavy chain variable domains derived from a repertoire of human heavy chain V, D, and J segments.

39. A method of generating an antibody that exhibits pH-dependent binding to an antigen of interest comprising:

generating a mouse of claim 22 ;

immunizing the mouse with an antigen of interest; and

selecting an antibody that binds to the antigen of interest with a desired affinity at a neutral pH while displaying reduced binding to the antigen of interest at an acidic pH.

40. The method of claim 39 , wherein the antibody exhibits a dissociative half-life (t 1/2 ) at acidic pH and 37° C. of about 2 minutes or less.

41. A method of making a non-human animal that comprises a genetically modified immunoglobulin light chain locus in its germline, the method comprising:

modifying a genome of a non-human animal to delete or render non-functional endogenous immunoglobulin light chain Vκ and Jκ segments in an immunoglobulin light chain locus, and

placing in the genome a single rearranged human light chain variable region gene sequence comprising human Vκ and Jκ segment sequences,

wherein the Vκ segment sequence is derived from a human Vκ1-39 or Vκ3-20 gene segment, and

wherein the single rearranged human immunoglobulin light chain variable region gene sequence comprises a substitution of at least one non-histidine codon of the Vκ segment sequence with a histidine codon that is expressed at a position selected from the group consisting of 105, 106, 107, 108, 109, 111 and a combination thereof (according to IMGT numbering).

42. The method of claim 41 , wherein the method results in a genetically modified non-human animal that comprises a population of B cells enriched for antibodies exhibiting pH-dependent binding to an antigen of interest.

43. The method of claim 41 , wherein the animal is a rodent.

44. The method of claim 43 , wherein the animal is a mouse or a rat.