Antigen-directed single domain antibody-based TNFR1 agonists elicit preferential killing of HER2-overexpressing cancer cells
Related Experiment Video
Updated: Apr 10, 2026
Analyzing Tumor and Tissue Distribution of Target Antigen Specific Therapeutic Antibody
Published on: May 16, 2020
Laura Unmuth1,2, Britta Lipinski1,2, Alicia Hoerr2
1Biomolecular Immunotherapy, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, 64287 Darmstadt, Germany.
Abstract:
In this study, we present a strategy to uncouple tumor necrosis factor (TNF)-like cell death induction from TNFR2 agonism in a tumor-targeted fashion. Single-domain antibodies (sdAbs) targeting TNFR1 were generated by combining camelid immunization with yeast surface display. Reformatting of resulting paratopes as bispecific antibodies (bsAbs) in a 2 + 2 manner by employing an sdAb-based paratope targeting HER2 revealed the identification of an immunocytokine-like bsAb, referred to as immunocytokine mimetic (ICM), which triggered TNF-like tumor cell death of HER2-overexpressing cancer cells as well as robust caspase-1, -3, and -8 activation in a cis-targeted manner. By modulating the valency of the TNFR1-directed sdAb, killing capacities as well as caspase activities of HER2-targeted ICMs were significantly augmented, eventually resulting in enhanced cell death induction when compared with TNF. Moreover, HER2-targeted TNFR1 ICMs also displayed a beneficial, i.e., substantially reduced profile in inducing unconditional pro-inflammatory cytokine release from peripheral blood mononuclear cells (PBMCs).
Insights
Researchers developed a novel bispecific antibody (bsAb) strategy to selectively induce tumor cell death via TNFR1 targeting. This approach minimizes harmful cytokine release, offering a safer cancer therapy option.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Tumor necrosis factor (TNF) induces cell death but lacks tumor specificity.
- Targeting TNF receptor 2 (TNFR2) can lead to unwanted inflammatory responses.
- Need for targeted strategies to induce TNF-like cell death specifically in tumors.
Purpose of the Study:
- To develop a method for uncoupling TNF-like cell death induction from TNFR2 agonism.
- To create a tumor-targeted bispecific antibody (bsAb) that selectively eliminates cancer cells.
- To assess the safety and efficacy of this novel therapeutic approach.
Main Methods:
- Generation of single-domain antibodies (sdAbs) targeting TNF receptor 1 (TNFR1).
- Reformatting sdAbs into a bispecific antibody (bsAb) format (2+2) with an HER2-targeting sdAb.
- Evaluation of bsAb-induced cell death, caspase activation, and cytokine release in cancer cells and PBMCs.
Main Results:
- Identified an immunocytokine mimetic (ICM) bsAb that triggers TNF-like cell death in HER2-overexpressing cancer cells.
- ICM demonstrated cis-targeted activation of caspases-1, -3, and -8, leading to enhanced tumor cell death.
- HER2-targeted TNFR1 ICMs showed reduced pro-inflammatory cytokine release from PBMCs compared to TNF.
Conclusions:
- The developed strategy successfully uncouples TNF-like cell death from TNFR2 agonism.
- HER2-targeted TNFR1 ICMs represent a promising approach for selective cancer cell killing.
- This method offers a potentially safer alternative to conventional TNF-based therapies with reduced systemic inflammation.
More Related Videos
16:07Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody mAb by Neutralizing TNF Using an In Vitro Bioanalytical Method
Published on: September 16, 2017
06:37Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety
Published on: September 15, 2023
Related Concept Videos
01:24Drugs for Treatment of Crohn's Disease in IBD Using Biologic Agents: Anti-TNF
01:27Tumor Immunotherapy