Human artificial chromosome containing human antibody λ light chain gene and non-human animal containing the human artificial chromosome capable of genetic transmission
View Patent ↗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.
1. A chimeric human antibody-producing mouse, wherein the antibody-producing mouse carries (1) human artificial chromosome ΔHAC or ΔΔHAC in which a 2.5 Mb region (ΔHAC) or a 1.5 Mb region (ΔΔHAC) from human chromosome 22 containing an unrearranged human antibody λ light chain locus is linked to a human chromosome 14 fragment containing an unrearranged human antibody heavy chain locus and (2) an unrearranged human antibody κ light chain transgene retained on KCo5 that is inserted into a mouse chromosome of the human antibody-producing mouse, at least both alleles of endogenous antibody heavy chain and κ light chain are disrupted or inactivated, and 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 are expressed in serum;
wherein at least 85% of the cells of the human antibody-producing mouse retain the human artificial chromosome ΔHAC or ΔΔHAC.
2. The human antibody-producing mouse according to claim 1 , which carries at least 40% of the variable region of the human antibody κ light chain.
3. The human antibody-producing mouse according to claim 1 , which carries all the variable regions of the human antibody heavy chain, the human antibody κ light chain, and the human antibody λ light chain.
4. The human antibody-producing mouse according to claim 1 , wherein the human antibody κ light chain locus inserted into the mouse chromosome is derived from human chromosome 2.
5. The human antibody-producing mouse according to claim 1 , wherein the human artificial chromosome is ΔΔHAC.
6. The human antibody-producing mouse according to claim 1 , wherein the human artificial chromosome is ΔHAC.
7. The human antibody-producing mouse according to claim 1 , wherein the human artificial chromosome is ΔΔHAC and 100% of the cells of the human antibody-producing mouse retain the human artificial chromosome.
8. A method for producing an antibody, which comprises: immunizing the mouse according to claim 1 , and obtaining a polyclonal antibody from the mouse.
9. The method according to claim 8 , wherein the polyclonal antibody is obtained from the blood of the mouse.
10. A method for producing an antibody, which comprises:
immunizing the mouse according to claim 1 with a desired antigen;
producing a hybridoma by fusing a mouse myeloma cell with a B cell derived from the mouse,
identifying the hybridoma that expresses a monoclonal antibody comprising human immunoglobulin heavy chains and light chains against the antigen, and producing the monoclonal antibody.