Humanized light chain mice
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsLynn Macdonald, Cagan Gurer, Karolina A. Hosiawa, Sean Stevens, Andrew J. Murphy
Life of this patent
Estimating the term…
Abstract
Non-human animals, tissues, cells, and genetic material are provided that comprise a modification of an endogenous non-human heavy chain immunoglobulin sequence and that comprise an ADAM6 activity functional in a mouse, wherein the non-human animals express a human immunoglobulin heavy chain variable domain and a cognate human immunoglobulin λ light chain variable domain.
Claims22
1-33. (canceled)
A method for generating a human heavy chain or human λ light chain variable region sequence comprising the steps of:
- (a) immunizing a genetically modified mouse with an antigen, wherein the genetically modified mouse:
- (i) has a germline genome that comprises:
- (1) an insertion of one or more human Vλ gene segments and one or more human Jλ gene segments upstream of an immunoglobulin light chain constant region gene,
- (2) an insertion of one or more human VH gene segments, one or more human DH gene segments, and one or more human JH gene segments upstream of a mouse immunoglobulin heavy chain constant region gene, and
- (3) an inserted nucleotide sequence that encodes a mouse ADAM6 protein, wherein the inserted nucleotide sequence that encodes a mouse ADAM6 protein is present at the immunoglobulin heavy chain locus of the genetically modified mouse, and wherein the mouse ADAM6 protein is expressed from the inserted nucleotide sequence;
- (ii) generates antibodies when immunized with the antigen, wherein the antibodies each comprise a human heavy chain variable domain operably linked to a mouse heavy chain constant domain and a human λ light chain variable domain operably linked to a light chain constant domain, and
- (iii) is fertile;
- and
- (b) identifying a human heavy chain or human λ light chain variable region sequence that encodes a human heavy chain or human λ light chain variable domain, respectively, of an antibody that specifically binds the antigen and that was generated by the genetically modified mouse.
The method of claim 43, wherein at least four human Jλ gene segments comprise at least Jλ1, Jλ2, Jλ3, and Jλ7.
A method of making a nucleotide sequence encoding a fully human heavy chain or a fully human light chain of an antibody that specifically binds an antigen comprising the steps of:
- (a) immunizing a genetically modified mouse with an antigen, wherein the genetically modified mouse:
- (i) has a germline genome that comprises:
- (1) an insertion of one or more human Vλ gene segments and one or more human Jλ gene segments upstream of an immunoglobulin light chain constant region gene,
- (2) an insertion of one or more human VH gene segments, one or more human DH gene segments, and one or more human JH gene segments upstream of a mouse immunoglobulin heavy chain constant region gene, and
- (3) an inserted nucleotide sequence that encodes a mouse ADAM6 protein, wherein the inserted nucleotide sequence is present at the immunoglobulin heavy chain locus of the genetically modified mouse, and wherein the mouse ADAM6 protein is expressed from the inserted nucleotide sequence,
- (ii) generates antibodies when immunized with the antigen, wherein the antibodies each comprise a human heavy chain variable domain operably linked to a mouse heavy chain constant domain and a human λ light chain variable domain operably linked to a light chain constant domain, and
- (iii) is fertile;
- and
- (b) identifying a human heavy chain or a human λ light chain variable region sequence that encodes a human heavy chain or human λ light chain variable domain. respectively, of an antibody that specifically binds the antigen and that was generated by the genetically modified mouse;
- and
- (c) operably linking the human heavy chain or the human λ light chain variable region sequence to a human heavy chain constant region sequence or a human light chain constant region sequence, respectively, to form a nucleotide sequence encoding fully human heavy chain or a fully human light chain.
A method of making genetically modified mouse, the method comprising introducing into its germline genome:
- (a) one or more human Vλ gene segments and one or more human Jλ gene segments upstream of an immunoglobulin light chain constant region gene,
- (b) one or more human VH gene segments, one or more human DH gene segments, and one or more human JH gene segments upstream of a mouse immunoglobulin heavy chain constant region, and
- (c) a nucleotide sequence that encodes a mouse ADAM6 protein, wherein the mouse ADAM6 protein is capable of restoring or enhancing fertility in a genetically modified male mouse.
Dependencies are inferred from the claim text.