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Tracking Bispecific Antibody-Induced T Cell Trafficking Using Luciferase-Transduced Human T Cells
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eIg-based bispecific T-cell engagers targeting EGFR: Format matters.

Lennart Kühl1, Annelie K Schäfer1, Sebastian Kraft1

  • 1Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.

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|March 2, 2023
PubMed
Summary

Bispecific T-cell engagers (TCEs) targeting EGFR and CD3 were engineered in various formats. Bivalent EGFR binding enhanced tumor cell killing, but TCE format critically impacts efficacy.

Keywords:
Bispecific antibodiesCD3EGFRT-cell retargetingvalency

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Area of Science:

  • Immunology
  • Biotechnology
  • Oncology

Background:

  • Bispecific antibodies, particularly T-cell engagers (TCEs), are promising cancer therapeutics.
  • TCEs redirect T-cells to tumor cells via dual antigen targeting (e.g., tumor antigen and CD3).
  • The molecular format and valency of TCEs can significantly influence their therapeutic activity.

Purpose of the Study:

  • To investigate how different molecular formats of bispecific TCEs targeting EGFR and CD3 affect their efficacy.
  • To evaluate the impact of Fc region presence, EGFR valency, and binding site arrangement on TCE function.

Main Methods:

  • Generation of 11 distinct bispecific TCE formats using the eIg platform, varying in Fc region inclusion and EGFR/CD3 binding site configuration (1+1 or 2+1).
  • Analysis of TCE binding to target and T-cells.
  • Assessment of T-cell-mediated tumor cell killing, cytokine release, and T-cell proliferation.

Main Results:

  • Bivalent binding to EGFR significantly enhanced TCE binding affinity and tumor cell killing.
  • The specific molecular composition and arrangement of the CD3-binding arm modulated target cell killing, cytokine release, and T-cell proliferation.
  • Different TCE formats exhibited varying levels of potency, highlighting format-dependent activity.

Conclusions:

  • Screening a diverse panel of bispecific TCE formats is crucial for identifying optimal candidates.
  • The molecular format of TCEs is a critical determinant of their therapeutic potency and T-cell activation profile.