Transgenic transchromosomal rodents for making human antibodies
View Patent ↗The present invention provides novel transgenic nonhuman mammals capable of producing human sequence antibodies, as well as methods of producing and using these antibodies.
1. A method for generating a plurality of B cells expressing human antibody sequences, the method comprising immunizing a transgenic mouse to generate a plurality of B cells expressing human antibody sequences, wherein the transgenic mouse comprises two human immunoglobulin loci, wherein one human immunoglobulin locus is a human heavy chain locus located on an autonomous transchromosome comprising a fragment of human chromosome 14 having a human chromosome 14 centromere, and the other human immunoglobulin locus is a human light chain locus transgene integrated into the mouse genome.
2. The method of claim 1 , further comprising isolating the plurality of B cells expressing human antibody sequences from the transgenic mouse.
3. The method of claim 2 , further comprising fusing the plurality of B cells with immortalized cells to form hybridomas.
4. The method of claim 3 , further comprising isolating the human antibody sequences from the hybridomas.
5. The method of claim 4 , wherein the human antibody sequences are purified.
6. The method of claim 1 , further comprising isolating nucleic acid sequences encoding the human antibody sequences from the transgenic mouse.
7. The method of claim 6 , wherein the nucleic acid sequences encoding the human antibody sequences are full length.
8. The method of claim 7 , further comprising expressing the nucleic acid sequences in a transfected cell.
9. The method of claim 1 , wherein the transchromosome comprising a fragment of human chromosome 14 is transchromosome SC20.
10. The method of claim 1 , wherein the transgene is KCo5 transgene.
11. A method for generating a human antibody that binds to a predetermined antigen, the method comprising:
immunizing a transgenic mouse with the predetermined antigen; and
isolating the human antibody from the immunized transgenic mouse,
wherein the transgenic mouse comprises two human immunoglobulin loci, wherein one human immunoglobulin locus is a human heavy chain locus located on an autonomous transchromosome comprising a fragment of human chromosome 14 having a human chromosome 14 centromere, and the other human immunoglobulin locus is a human light chain locus transgene integrated into the mouse genome.
12. The method of claim 11 , wherein the human antibody binds to a predetermined antigen with an equilibrium association constant (Ka) of at least 10 10 M −1 .
13. The method of claim 11 , wherein the human antibody is an IgG isotype or an IgA isotype.
14. The method of claim 11 , wherein the human antibody is antigen-specific.
15. A method for generating antigen-specific hybridomas secreting a human antibody, the method comprising:
immunizing a transgenic mouse with a predetermined antigen;
fusing lymphocytes from the transgenic mouse with immortalized cells to form hybridoma cells; and
identifying hybridomas that secrete a human antibody that specifically binds to the predetermined antigen,
wherein the transgenic mouse comprises two human immunoglobulin loci, wherein one human immunoglobulin locus is a human heavy chain locus located on an autonomous transchromosome comprising a fragment of human chromosome 14 having a human chromosome 14 centromere, and the other human immunoglobulin locus is a human light chain locus transgene integrated into the mouse genome.
16. A method for generating a human antibody that binds to a predetermined antigen, the method comprising:
immunizing a transgenic mouse with a predetermined antigen;
fusing lymphocytes from the transgenic mouse with immortalized cells to form hybridoma cells; and
screening hybridoma cells formed for the presence of antigen reactive antibodies, wherein the transgenic mouse comprises two human immunoglobulin loci, wherein one human immunoglobulin locus is a human heavy chain locus located on an autonomous transchromosome comprising a fragment of human chromosome 14 having a human chromosome 14 centromere, and the other human immunoglobulin locus is a human light chain locus transgene integrated into the mouse genome.
17. The method of claim 16 , wherein the antigen reactive antibodies are secreted from the hybridoma in culture.