Construction and application of bispecific antibody EpCAM×CD3
The present invention provides a bispecific antibody. The bispecific antibody provided by the present invention comprises a single-chain unit and a monovalent unit, wherein the single-chain unit has a specific binding capability against surface antigen CD3 of an immune cell; the monovalent unit has a specific binding capability against the surface antigen EpCAM of a tumor cell; the single-chain unit comprises a single-chain variable fragment ScFv fused with an Fc fragment; and the monovalent unit comprises a light chain and heavy chain pair. The present invention also provides a preparation method of the bispecific antibody and pharmaceutical use of these antibodies.
1. A bispecific antibody, comprising:
(a) a monovalent unit comprising a light chain-heavy chain pair which specifically binds to EpCAM, wherein the heavy chain comprises the amino acid sequence of SEQ ID NO: 1 and the light chain comprises the amino acid sequence of SEQ ID NO: 3: and
(b) a single-chain unit which is a fusion peptide comprising a single-chain variable fragment (scFv) and an FC fragment which comprises a hinge region, a CH2 structural domain and a CH3 structural domain, wherein the single-chain unit has binding specificity to a surface antigen CD3, and wherein the single-chain unit comprises the amino acid sequence of SEQ ID NO: 5.
2. The bispecific antibody of claim 1 , wherein the CH2 structural domain of the single-chain unit is positioned between the scFv fragment and the CH3 structural domain and the single-chain unit does not contain a CH1 structural domain.