title: Tetravalent anti-CD20/CD3 bispecific antibody for the treatment of B cell lymphoma.
abstract: Bispecific antibodies (bsAbs) are second generation antibodies for therapeutic application in immunotherapy. One of the major strategies of the bsAb platform is the recruitment of immune effector T cells by incorporating an anti-CD3 domain. A bispecific T-cell engager (BiTE), with one end having an affinity for CD3 and the other end with affinity for CD19, has been approved in the US and Europe for the treatment of acute lymphoblastic leukemia. However, due to their small size and lack of Fc region, these single-chain variable fragment (scFv) bsAbs have short half-lives in vivo. Additionally, poor solubility, structural instability, and low production yields have also become major challenges in the bulk production process. To overcome these challenges, we have engineered a tetravalent bsAb with bivalent binding specificity for the CD20 and CD3 antigen in an immunoglobulin G (IgG) format. The fusion of the anti-CD3 scFvs to the CD20 antibody via a linker-hinge domain (LHD) results in improved antibody stabilization and properties. Here we demonstrate this antibody's highly efficient cancer cell elimination in a dose-dependent manner in a CD20-expressing B lymphoblastoid cell line in vitro. Our data suggest the potential clinical application of this bsAb for the treatment of CD20-expressing B cell malignancies.
authors.affiliation: Institute of Biologics, Development Center for Biotechnology, New Taipei City, Taiwan.
authors.forename: Chia-Yen
authors.initials: CY
authors.lastname: Lu
authors.affiliation: Institute of Biologics, Development Center for Biotechnology, New Taipei City, Taiwan.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Chen
authors.affiliation: Institute of Biologics, Development Center for Biotechnology, New Taipei City, Taiwan.
authors.forename: Pei-Han
authors.initials: PH
authors.lastname: Tai
authors.affiliation: Institute of Biologics, Development Center for Biotechnology, New Taipei City, Taiwan.
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Yang
authors.affiliation: Laboratory of Biopharmaceutical Research, Advagene Biopharma, Taipei, Taiwan. Electronic address: yshsu@advagene.com.tw.
authors.forename: Yu-Shen
authors.initials: YS
authors.lastname: Hsu
authors.affiliation: Laboratory of Biopharmaceutical Research, Advagene Biopharma, Taipei, Taiwan. Electronic address: mingi.chang@advagene.com.tw.
authors.forename: Mingi
authors.initials: M
authors.lastname: Chang
authors.affiliation: Institute of Biologics, Development Center for Biotechnology, New Taipei City, Taiwan. Electronic address: fabio@dcb.org.tw.
authors.forename: Chuan-Lung
authors.initials: CL
authors.lastname: Hsu
mesh_terms: D000911:Antibodies, Monoclonal / Q000008:administration & dosage* / Q000276:immunology / Q000494:pharmacology; D018951:Antigens, CD20 / Q000276:immunology*; D017209:Apoptosis / Q000187:drug effects; D017252:CD3 Complex / Q000276:immunology*; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004355:Drug Stability; D006801:Humans; D019169:Jurkat Cells; D016393:Lymphoma, B-Cell / Q000188:drug therapy* / Q000276:immunology* / Q000473:pathology; D015202:Protein Engineering; D016896:Treatment Outcome
chemical_list: D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; D017252:CD3 Complex
keywords: Anti-CD3; Bispecific T-cell engager; Bispecific antibody; CD20