Shortened hinge design of Fab x sdAb-Fc bispecific antibodies enhances redirected T-Cell killing of tumor cells

Shuyu Huang, Aina Segués, Martin Waterfall, David Wright, Charlotte Vayssiere, Sander M. J. Van Duijnhoven, Andrea Van Elsas, Alice J. A. M. Sijts, Dietmar M. Zaiss

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

T cell engager (TCE) antibodies have emerged as promising cancer therapeutics that link cytotoxic T-cells to tumor cells by simultaneously binding to CD3E on T-cells and to a tumor-associated antigen (TAA) expressed by tumor cells. We previously reported a novel bispecific format, the IgG-like Fab x sdAb-Fc (also known as half-IG_VH-h-CH2-CH3), combining a conventional antigen-binding fragment (Fab) with a single domain antibody (sdAb). Here, we evaluated this Fab x sdAb-Fc format as a T-cell redirecting bispecific antibody (TbsAbs) by targeting mEGFR on tumor cells and mCD3E on T cells. We focused our attention specifically on the hinge design of the sdAb arm of the bispecific antibody. Our data show that a TbsAb with a shorter hinge of 23 amino acids (TbsAb.short) showed a significantly better T cell redirected tumor cell elimination than the TbsAb with a longer, classical antibody hinge of 39 amino acids (TbsAb.long). Moreover, the TbsAb.short form mediated better T cell-tumor cell aggregation and increased CD69 and CD25 expression levels on T cells more than the TbsAb.long form. Taken together, our results indicate that already minor changes in the hinge design of TbsAbs can have significant impact on the anti-tumor activity of TbsAbs and may provide a new means to improve their potency.
Original languageEnglish
Article number1331
Number of pages16
Issue number10
Publication statusPublished - 21 Sept 2022

Keywords / Materials (for Non-textual outputs)

  • bispecific antibody
  • cancer immunotherapy
  • mCD3E
  • mEGFR
  • hinge


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