Genetic engineering of non-human animals for the production of chimeric antibodies
The invention provides non-human cells and mammals having a genome encoding chimeric antibodies and methods of producing transgenic cells and mammals. Certain aspects of the invention include chimeric antibodies, humanized antibodies, pharmaceutical compositions and kits. Certain aspects of the invention also relate to diagnostic and treatment methods using the antibodies of the invention.
1. An isolated mammalian cell whose genome comprises a transgene comprising (1) a plurality of immunoglobulin light chain variable (V) exons encoding human immunoglobulin light chain variable (V) polypeptides; (2) non-coding sequences between the V exons; (3) a plurality of immunoglobulin light chain joining (J) coding sequences encoding human immunoglobulin light chain joining (J) polypeptides; and (4) non-coding sequences between the J coding sequences; wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are derived from mouse, rat, or a combination thereof immunoglobulin light chain non-coding sequences.
2. The isolated mammalian cell according to claim 1 , wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are selected from the group consisting of intronic sequences and cis regulatory sequences.
3. The isolated mammalian cell according to claim 2 , wherein the cis regulatory sequences are selected from promoters, enhancers, recombination signal sequences, splice acceptor sequences, and splice donor sequences.
4. The isolated mammalian cell according to claim 1 , wherein the V exons encode human kappa light chain V (Vκ) polypeptides or human lambda light chain V (Vλ) polypeptides.
5. The isolated mammalian cell according to claim 4 , wherein the transgene is capable of undergoing rearrangement to produce a polynucleotide sequence encoding a polypeptide comprising an immunoglobulin light chain variable region.
6. The isolated mammalian cell according to claim 1 , wherein (1) the non-coding sequences between the V exons and (2) the V exons are from non-orthologous immunoglobulin loci.
7. The isolated mammalian cell according to claim 1 , wherein the transgene further comprises a coding sequence encoding a human immunoglobulin light chain constant (C) polypeptide.
8. The isolated mammalian cell according to claim 7 , wherein the coding sequence encoding the human immunoglobulin light chain C polypeptide encodes a human Cλ polypeptide or a human Cκ polypeptide.
9. The isolated mammalian cell according to claim 1 , wherein the cell is murine.
10. The isolated mammalian cell according to claim 1 , wherein the cell is human.
11. The isolated mammalian cell according to claim 1 , wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are mouse immunoglobulin light chain non-coding sequences.
12. The isolated mammalian cell according to claim 1 , wherein the non-coding sequences between the V exons and the non-coding sequences between the J coding sequences are rat immunoglobulin light chain non-coding sequences.
13. The isolated mammalian cell according to claim 2 , wherein the cis regulatory sequences are endogenous.
14. The isolated mammalian cell according to claim 1 , wherein the transgene further comprises a coding sequence encoding a mouse immunoglobulin light chain constant (C) polypeptide.
15. The isolated mammalian cell according to claim 14 , wherein the transgene comprises human Vλ exons and 2 to 7 Jλ-Cλ coding sequence pairs, wherein the 2 to 7 Jλ sequences are human Jλ and the 2 to 7 Cλ sequences are mouse Cλ.
16. The isolated mammalian cell according to claim 1 , wherein the transgene further comprises mouse, rat, or a combination thereof non-coding sequences upstream of the V exons.
17. The isolated mammalian cell according to claim 1 , wherein the transgene further comprises mouse, rat, or a combination thereof non-coding sequences downstream of the J coding sequences.
18. An isolated mammalian cell whose genome comprises a transgene comprising (1) a plurality of human immunoglobulin light chain variable Vλ exons encoding human immunoglobulin light chain variable Vλ polypeptides; (2) mouse non-coding sequences between the Vλ exons; (3) 2-7 immunoglobulin light chain Jλ-Cλ coding sequence pairs, wherein the Jλ sequence encodes a human immunoglobulin light chain Jλ polypeptide and the Cλ sequence encodes a mouse immunoglobulin light chain Cλ polypeptide; (4) mouse non-coding sequences between the Jλ coding sequences and the Cλ, coding sequences in the Jλ-Cλ coding sequence pairs; and (5) mouse non-coding sequences between the Jλ-Cλ coding sequence pairs.
19. The isolated mammalian cell according to claim 18 , wherein the transgene comprises 6 Jλ-Cλ coding sequence pairs.
20. The isolated mammalian cell according to claim 18 , wherein the transgene further comprises mouse non-coding sequences upstream of the Vλ exons.
21. The isolated mammalian cell according to claim 18 , wherein the transgene further comprises mouse non-coding sequences downstream of the Jλ-Cλ coding sequence pairs.
22. An isolated mammalian cell whose genome comprises a transgene comprising (1) a plurality of human immunoglobulin light chain variable Vκ exons encoding human immunoglobulin light chain variable Vκ polypeptides; (2) mouse non-coding sequences between the Vκ exons; (3) a plurality of human immunoglobulin light chain variable Jκ coding sequences encoding human immunoglobulin light chain variable Jκ polypeptides; (4) mouse non-coding sequences between the Jκ coding sequences; and (5) a Cκ coding sequence encoding a mouse immunoglobulin light chain constant Cκ polypeptide.
23. The isolated mammalian cell according to claim 22 , wherein the transgene further comprises mouse non-coding sequences upstream of the Vκ exons.
24. The isolated mammalian cell according to claim 22 , wherein the transgene further comprises mouse non-coding sequences downstream of the Cκ coding sequence.
25. The isolated mammalian cell according to claim 22 , wherein the transgene further comprises 5 Jκ exons.
26. The isolated mammalian cell according to claim 18 , wherein the transgene further comprises a human immunoglobulin light chain λ 3′ enhancer.
27. The isolated mammalian cell according to claim 18 , wherein said transgene further comprises an Ig light chain 3′LCR, or a functional fragment thereof.
28. The isolated mammalian cell according to claim 27 , wherein said Ig light chain 3′LCR, or a functional fragment thereof, is from a mammal selected from the group consisting of human, non-primate and rat.