Transgenic animals producing monovalent human antibodies and antibodies obtainable from these animals
AssigneeGenmab A/S
InventorsJanine Schuurman, Tom Vink, Jan Van De Winkel, Aran Frank Labrijn, Paul Parren, Willem Karel Bleeker, Frank Beurskens, Patrick Van Berkel
Life of this patent
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Abstract
The invention relates to novel non-human transgenic animals, which upon antigenic stimulation are capable of producing monovalent antibodies binding to a selected antigen, modified heavy chain transgenes, methods for producing the non-human transgenic animals, methods for immunizing the non-human transgenic animals for as well as monovalent antibodies obtainable by such immunization methods.
Claims73
A non-human transgenic animal, which upon antigenic stimulation is capable of producing a monovalent antibody that binds to a selected antigen, which monovalent antibody comprises a heavy chain, which heavy chain comprises (i) a human VH region, and (ii) a human CH region, wherein the CH region has been modified such that the region corresponding to the hinge region and, optionally other regions of the C H region, such as the CH3 region, do not comprise any amino acid residues which are capable of forming disulfide bonds with an identical C H region or other covalent or stable non-covalent inter-heavy chain bonds with an identical CH region in the presence of polyclonal human IgG.
A non-human transgenic animal, which upon antigenic stimulation is capable of producing a monovalent IgG4 antibody that binds to a selected antigen, which monovalent antibody comprises
- a heavy chain, which heavy chain comprises (i) a human VH region, and (ii) a human CH region, wherein the CH region has been modified such that the region corresponding to the hinge region does not comprise any amino acid residues which are capable of forming disulfide bonds with an identical CH region, and
- a light chain, which light chain comprises (i) a human VL region, and (ii) a human CL region.
A non-human transgenic animal, which upon antigenic stimulation is capable of producing a monovalent IgG4 antibody that binds to a selected antigen, which monovalent antibody comprises
- a heavy chain, which heavy chain comprises
- (i) a human VH region, and
- (ii) a human CH region, wherein the CH region has been modified such that the region corresponding to the hinge region does not comprise any amino acid residues which are capable of forming disulfide bonds with an identical CH region, and wherein the CH3 region has the sequence as set forth in SEQ ID NO:
- 16, but wherein the CH3 region has been modified so that one or more of the following amino acid substitutions have been made:
- Thr (T) in position 234 has been replaced by Ala (A);
- Leu (L) in position 236 has been replaced by Ala (A);
- Leu (L) in position 236 has been replaced by Val (V);
- Phe (F) in position 273 has been replaced by Ala (A);
- Phe (F) in position 273 has been replaced by Leu (L);
- Tyr (Y) in position 275 has been replaced by Ala (A);
- Arg (R) in position 277 has been replaced by Ala (A), and
- a light chain, which light chain comprises
- (i) a human VL region, and
- (ii) a human CL region.
A non-human transgenic animal, which upon antigenic stimulation is capable of producing a monovalent IgG1 antibody that binds to a selected antigen, which monovalent antibody comprises a heavy chain, which heavy chain comprises (i) a human VH region, and (ii) a human CH region, wherein the CH region has been modified such that the regions corresponding to the hinge region and the CH3 region, do not comprise any amino acid residues which are capable of forming disulfide bonds with an identical CH region or other covalent or stable non-covalent inter-heavy chain bonds with an identical CH region in the presence of polyclonal human IgG, and a light chain, which light chain comprises (i) a human VL region, and (ii) a human CL region, which has been modified such that the CL region does not contain any amino acids which are capable of forming disulfide bonds with an identical CL region or other covalent bonds with an identical CL region in the presence of polyclonal human IgG.
A heavy chain transgene comprising (i) a plurality of human V genes, a plurality of human D genes, a plurality of human J genes, and (ii) a plurality of human CH genes and associated isotype switch sequences, comprising a human μ CH gene and at least one modified γ CH gene, wherein the human γ CH gene is in closer proximity to the human μ CH gene than in a naturally occurring human immunoglobulin heavy chain gene locus.
A monovalent IgG4 antibody that binds to a selected antigen, which monovalent antibody comprises
- a heavy chain, which heavy chain comprises
- (i) a human VH region, and
- (ii) a human CH region, wherein the CH region has been modified such that the region corresponding to the hinge region does not comprise any amino acid residues which are capable of forming disulfide bonds with an identical CH region, and wherein the CH3 region has the sequence as set forth in SEQ ID NO:
- 16, but wherein the CH3 region has been modified so that one or more of the following amino acid substitutions have been made:
- Thr (T) in position 234 has been replaced by Ala (A);
- Leu (L) in position 236 has been replaced by Ala (A);
- Leu (L) in position 236 has been replaced by Val (V);
- Phe (F) in position 273 has been replaced by Ala (A);
- Phe (F) in position 273 has been replaced by Leu (L);
- Tyr (Y) in position 275 has been replaced by Ala (A);
- Arg (R) in position 277 has been replaced by Ala (A), and
- a light chain, which light chain comprises
- (i) a human VL region, and
- (ii) a human CL region.
The monovalent antibody according to aim 49, wherein the monovalent antibody further comprises the CH2 region as set forth in SEQ ID NO:
- 16, but wherein Asn (N) in position 302 has been replaced by Ala (A) and Thr (T) in position 175 has been replaced by Ala (A) and Glu (E) in position 248 has been replaced by Ala (A).
Dependencies are inferred from the claim text.