Bispecific antibody
Provided are bispecific antibodies comprised of a single-chain unit having specificity to an immune cell and a monovalent unit having specificity to a tumor cell or a microorganism. The single-chain unit includes a single-chain variable fragment (scFv) fused to an Fc fragment and the monovalent unit includes a light chain and heavy chain pair. Also provided are methods of preparing bispecific antibodies and pharmaceutical and diagnostic uses of these antibodies.
1. An antibody comprising:
(a) a monovalent unit comprising a light chain-heavy chain pair having specificity to Her2, wherein the light chain of the monovalent unit comprises SEQ ID NO: 5, and wherein the heavy chain of the monovalent unit comprises SEQ ID NO: 3; and
(b) a single-chain unit comprising a fusion peptide comprising a single-chain variable fragment (scFv) having specificity to CD3 and an Fc fragment comprising a CH2 domain and a CH3 domain, wherein the fusion peptide comprises SEQ ID NO: 1.
2. The antibody of claim 1 , wherein the light chain is bound to the heavy chain through a disulfide bond.
3. The antibody of claim 1 , wherein the heavy chain is bound to the fusion peptide through one or more disulfide bonds.
4. The antibody of claim 1 , wherein the antibody is a humanized antibody.
5. The antibody of claim 1 , further comprising a detectable label.
6. The antibody of claim 1 , wherein the antibody binds Her2 expressing breast cancer cells and one or more CD3 expressing cells selected from the group consisting of a lymphocyte, a monocyte and a macrophage.
7. The antibody of claim 1 , wherein the Fc region of one unit has the following amino acid substitutions: Threonine (T) to Tryptophan (W) substitution at position 366, Lysine (K) to Aspartic acid (D) substitution at position 392, and Lysine (K) to Aspartic acid (D) substitution at position 409 (Kabat numbering system); and
wherein the Fc region of the other unit has the following amino acid substitutions: Leucine (L) to Arginine (R) substitution at position 368, Aspartic acid (D) to Lysine (K) substitution at position 399, and Tyrosine (Y) to Alanine (A) substitution at position 407 (Kabat numbering system).
8. The antibody of claim 1 , wherein the Fc region of the heavy chain of the monovalent unit has the following amino acid substitutions: Threonine (T) to Tryptophan (W) substitution at position 366, Lysine (K) to Aspartic acid (D) substitution at position 392, and Lysine (K) to Aspartic acid (D) substitution at position 409 (Kabat numbering system); and
wherein the Fc region of the fusion peptide has the following amino acid substitutions: Leucine (L) to Arginine (R) substitution at position 368, Aspartic acid (D) to Lysine (K) substitution at position 399, and Tyrosine (Y) to Alanine (A) substitution at position 407 (Kabat numbering system).