Human artificial chromosome containing human antibody λ light chain gene and non-human animal containing the human artificial chromosome capable of genetic transmission

Patent metadata

Field	Value
Publication number	US-9499838-B2
Application number	US-14000908-A
Country	US
Kind code	B2
Filing date	Jun 16, 2008
Priority date	May 11, 2001
Publication date	Nov 22, 2016
Grant date	Nov 22, 2016

Abstract

The present invention relates to a human artificial chromosome which is genetically transmissible to the next generation with high efficiency and the method for using the same. More specifically, the present invention relates to: a human artificial chromosome in which an about 3.5 Mb to about 1 Mb region containing an antibody λ light chain gene derived from human chromosome 22 is bound to a chromosome fragment which is transmissible to a progeny through a germ line of a non-human animal, said chromosome fragment is derived from another human chromosome; a non-human animal carrying the human artificial chromosome and an offspring thereof; a method for producing the non-human animal; a method for producing a human antibody using the nonhuman animal or an offspring thereof; and a human antibody-producing mouse carrying the human artificial chromosome.

First claim

The invention claimed is:

1. A method for producing a monoclonal antibody, which comprises:
immunizing with a desired antigen, a human antibody-producing mouse which carries (1) a human artificial chromosome in which a SC20 chromosome vector is linked to an about 1 Mb to about 3.5 Mb fragment of human chromosome 22 comprising an unrearranged human antibody λ light chain locus, and (2) an unrearranged human antibody κ light chain locus retained on a KCo5 transgene or on fragment W23 of human chromosome 2 (hCF(2-W23)), wherein at least both alleles of endogenous heavy chain locus and κ light chain locus are disrupted or inactivated, and which expresses a human antibody heavy chain comprising a human antibody Ig γ isotype, a human antibody κ light chain, and a human antibody λ light chain;
producing a hybridoma by fusing a mouse myeloma cell with the a spleen cell derived from the mouse; and
thereby producing a monoclonal antibody comprising human immunoglobulin heavy chains and light chains against the antigen.

2. The method of claim 1 , wherein the unrearranged human antibody κ light chain locus is inserted into a mouse chromosome.

3. The method of claim 1 , which further comprises:
isolating human heavy and light chain antibody genes from the hybridoma;
introducing the human heavy and light chain antibody genes into a cell capable of expressing the human heavy and light chain antibody genes;
culturing the cell under a condition capable of expressing the human heavy and light chain antibody genes; and
thereby producing a monoclonal antibody comprising heavy chains and light chains.

4. The method according to claim 3 , wherein the cell is an animal cell, a yeast cell or an insect cell.

5. The method of claim 1 , wherein the human antibody-producing mouse carries at least 40% of the variable region of the human antibody κ light chain.

6. The method of claim 1 , wherein the human antibody-producing mouse carries all the variable regions of the human antibody heavy chain, the human antibody κ light chain, and the human antibody λ light chain.

7. The method of claim 1 , wherein the human antibody κ light chain locus is retained on hCF(2-W23).

8. The method of claim 1 , wherein the human artificial chromosome is ΔΔHAC.

9. The method of claim 1 , wherein the human artificial chromosome is ΔHAC.

10. The method of claim 8 , wherein 100% of cells of the human antibody-producing mouse retain the human artificial chromosome.

11. The method of claim 1 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is from about 1.2 Mb to about 3 Mb.

12. The method of claim 1 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is from about 1.5 Mb to about 2.5 Mb.

13. The method of claim 1 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is 1.5 Mb.

14. The method of claim 1 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is 2.5 Mb.

15. A method for producing a monoclonal antibody, which comprises:
immunizing with a desired antigen, a human antibody-producing mouse which carries a human artificial chromosome in which a SC20 chromosome vector is linked to an about 1 Mb to about 3.5 Mb fragment of human chromosome 22 comprising an unrearranged human antibody λ light chain locus, wherein at least both alleles of endogenous heavy chain locus and κ light chain locus are disrupted or inactivated, and which expresses a human antibody heavy chain comprising a human antibody Ig γ isotype and a human antibody λ light chain;
producing a hybridoma by fusing a mouse myeloma cell with the a spleen cell derived from the mouse; and
thereby producing a monoclonal antibody comprising human immunoglobulin heavy chains and light chains against the antigen.

16. The method of claim 15 , which further comprises:
isolating human heavy and light chain antibody genes from the hybridoma;
introducing the human heavy and light chain antibody genes into a cell capable of expressing the human heavy and light chain antibody genes;
culturing the cell under a condition capable of expressing the human heavy and light chain antibody genes; and
thereby producing a monoclonal antibody comprising heavy chains and light chains.

17. The method according to claim 16 , wherein the cell is an animal cell, a yeast cell or an insect cell.

18. The method of claim 15 , wherein the human artificial chromosome is ΔΔHAC.

19. The method of claim 15 , wherein the human artificial chromosome is ΔHAC.

20. The method of claim 18 , wherein 100% of cells of the human antibody-producing mouse retain the human artificial chromosome.

21. The method of claim 15 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is from about 1.2 Mb to about 3 Mb.

22. The method of claim 15 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is from about 1.5 Mb to about 2.5 Mb.

23. The method of claim 15 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is 1.5 Mb.

24. The method of claim 15 , wherein the size of the fragment of human chromosome 22 comprising the unrearranged human antibody λ light chain locus is 2.5 Mb.

Assignees

• Kuroiwa Yoshimi
• Tomizuka Kazuma
• Yoshida Hitoshi
• Ishida Isao
• Kyowa Hakko Kirin Co Ltd
• Squibb & Sons Llc

Inventors

• Kuroiwa Yoshimi
• Tomizuka Kazuma
• Yoshida Hitoshi
• Ishida Isao

Classifications

• C12N15/90

  Stable introduction of foreign DNA into chromosome · CPC title

• A01K2207/15

  Humanized animals · CPC title

• A61K48/00

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title

• C07K16/243

  Colony Stimulating Factors · CPC title

• A01K2217/054

  inducing loss of function · CPC title