Humanized universal light chain mice

A method for generating a human immunoglobulin heavy chain variable domain sequence of an antibody that specifically binds to an antigen of interest comprising the steps of:

• (a) immunizing a genetically modified mouse with an antigen of interest, wherein the mouse:
• (i) has a germline genome that comprises:
• (A) an insertion comprising at least one unrearranged human VH gene segment, at least one unrearranged human DH gene segment, and at least one unrearranged human JH gene segment, wherein the at least one unrearranged human VH gene segment, at least one unrearranged human DH gene segment, and at least one unrearranged human JH gene segment are operably linked to a heavy chain constant region gene, wherein the insertion disrupts the function of an endogenous ADAM6 protein, and wherein the disruption of the endogenous ADAM6 function is associated with a reduction in fertility in male mice;
• (B) an insertion comprising a single rearranged light chain V/J sequence, wherein the single rearranged light chain V/J sequence is operably linked to a light chain constant region gene, wherein the single rearranged light chain V/J sequence comprises a human Vκ1-39;
• and
• (C) an insertion comprising an ectopic nucleic acid sequence that encodes a functional mouse ADAM6 protein, which nucleic acid sequence is integrated in the germline genome of the mouse,
• wherein, when the mouse is a male, the functional mouse ADAM6 protein is expressed and the male mouse has wild type fertility;
• (ii) generates antibodies when immunized with the antigen of interest, wherein the antibodies each comprise a human heavy chain variable domain operably linked to a heavy chain constant domain and a light chain variable domain operably linked to a light chain constant domain, wherein the light chain variable domain is expressed from the rearranged light chain V/J sequence in the germline genome of the mouse or from a somatically hypermutated variant thereof;
• and
• (b) determining a human heavy chain variable domain sequence of an antibody that specifically binds the antigen of interest and that was generated by the genetically modified mouse.

A method of making an antibody comprising:

• (a) immunizing a genetically modified mouse with an antigen of interest, wherein the mouse:
• (i) has a germline genome that comprises:
• (A) an insertion comprising at least one unrearranged human VH gene segment, at least one unrearranged human DH gene segment, and at least one unrearranged human JH gene segment, wherein the at least one unrearranged human VH gene segment, at least one unrearranged human DH gene segment, and at least one unrearranged human JH gene segment are operably linked to a heavy chain constant region gene, wherein the insertion disrupts the function of an endogenous ADAM6 protein, and wherein the disruption of the endogenous ADAM6 function is associated with a reduction in fertility in male mice;
• (B) an insertion comprising a single rearranged light chain V/J sequence, wherein the single rearranged light chain V/J sequence is operably linked to a light chain constant region gene, wherein the single rearranged light chain V/J sequence comprises a human Vκ1-39;
• and
• (C) an insertion comprising an ectopic nucleic acid sequence that encodes a functional mouse ADAM6 protein, which nucleic acid sequence is integrated in the germline genome of the mouse,
• wherein, when the mouse is a male, the functional mouse ADAM6 protein is expressed and the male mouse has wild type fertility;
• (ii) generates antibodies when immunized with the antigen of interest, wherein the antibodies each comprise a human heavy chain variable domain operably linked to a heavy chain constant domain and a light chain variable domain operably linked to a light chain constant domain, wherein the light chain variable domain is expressed from the single rearranged light chain V/J sequence in the germline genome of the mouse or from a somatically hypermutated variant thereof;
• and
• (b) determining an amino acid sequence of a human variable domain of the antibody that specifically binds the antigen of interest or determining a nucleotide sequence that encodes a human variable domain of the antibody that specifically binds the antigen of interest;
• and
• (c) employing the amino acid sequence or the nucleotide sequence to produce the antibody.