Antibody producing non-human mammals
Described are transgenic, non-human animals comprising a nucleic acid encoding an immunoglobulin light chain, whereby the immunoglobulin light chain is human, human-like, or humanized. The nucleic acid is provided with a means that renders it resistant to DNA rearrangements and/or somatic hypermutations. In one embodiment, the nucleic acid comprises an expression cassette for the expression of a desired molecule in cells during a certain stage of development in cells developing into mature B cells. Further provided is methods for producing an immunoglobulin from the transgenic, non-human animal.
1. A transgenic mouse that has been immunized with an antigen wherein the genome of said mouse comprises a transgene comprising a single human immunoglobulin light chain germline V gene segment fused to a single human immunoglobulin light chain germline J gene segment such that there is no mutation due to said fusion,
wherein said fused human V/J gene segments encode a rearranged human immunoglobulin light chain variable region, wherein the transgene is inserted by site-specific integration in the murine Rosa locus or wherein said transgene lacks the intronic light chain enhancer MoEκi or comprises a truncation of the transgene 3′ kappa enhancer or combination of these; and
wherein the transgene comprises a murine light chain constant region gene segment or is operatively linked to an endogenous mouse light chain constant region gene segment;
wherein if the transgene is inserted by site-specific integration in the murine Rosa locus the transgene comprises a murine light chain constant region gene segment; and
wherein said transgenic mouse, in response to said antigen, has produced B cells that secrete antibodies with immunoglobulin light chains comprising said rearranged human light chain variable region and a murine light chain constant region, paired with a diversity of immunoglobulin heavy chains, which bind said antigen.
2. A method to prepare antibodies immunoreactive with a desired antigen which method comprises isolating B cells from the transgenic mouse of claim 1 and culturing said cells, optionally immortalizing said cells and harvesting the antibodies.
3. A method to prepare antibodies immunoreactive with a desired antigen which method comprises isolating nucleic acids from the B cells of the transgenic mouse of claim 1 that encode at least part of such antibodies, inserting the nucleic acids or copies or derivatives thereof into an expression cassette and expressing the antibodies in host cells.
4. A method to prepare antibodies immunoreactive with a desired antigen which method comprises recovering said antibodies from the mouse of claim 1 .