Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors

Abstract Fusion proteins combining TNF-related apoptosis inducing ligand (TRAIL) and antibody building blocks have emerged as a strategy for the targeted treatment of cancer cells. Using a single-chain derivative of homotrimeric TRAIL (scTRAIL), several targeted and non-targeted scTRAIL fusion prote...

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Main Authors: Dennis Michler, Oliver Seifert, Klaus Pfizenmaier, Roland E. Kontermann
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-00476-7
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author Dennis Michler
Oliver Seifert
Klaus Pfizenmaier
Roland E. Kontermann
author_facet Dennis Michler
Oliver Seifert
Klaus Pfizenmaier
Roland E. Kontermann
author_sort Dennis Michler
collection DOAJ
description Abstract Fusion proteins combining TNF-related apoptosis inducing ligand (TRAIL) and antibody building blocks have emerged as a strategy for the targeted treatment of cancer cells. Using a single-chain derivative of homotrimeric TRAIL (scTRAIL), several targeted and non-targeted scTRAIL fusion proteins of varying geometries and valencies for TRAIL receptors and target antigens, all comprising an Fc region, were generated. These fusion proteins comprised either 1 or 2 scTRAIL units, i.e. are tri- or hexavalent for TRAIL receptors and in the targeted versions, 1 or 2 binding sites for EGFR. These fusion proteins were analyzed for cell binding and cell death induction using the EGFR-expressing colorectal cancer cell lines Colo205 and HCT116. In line with previous findings, all fusion proteins that were hexavalent for TRAIL receptors exhibited a strongly increased cell killing activity compared to the trivalent ones. Interestingly, the fusion proteins comprising one scTRAIL unit, did not benefit from targeting to EGFR. In contrast, the hexavalent scTRAIL fusion proteins further benefited from EGFR targeting, resulting in an approximately 6- to 30-fold increase in cell killing. In summary, this study shed further light on the influence of geometry and valency of TRAIL fusion proteins and confirmed IgG-scTRAIL fusion proteins as highly potent cell death inducers.
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spelling doaj-art-361e15ecada94dcaa9ff7fdcfd0ec0ef2025-08-20T03:09:35ZengNature PortfolioScientific Reports2045-23222025-05-0115111010.1038/s41598-025-00476-7Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptorsDennis Michler0Oliver Seifert1Klaus Pfizenmaier2Roland E. Kontermann3Institute of Cell Biology and Immunology, University of StuttgartInstitute of Cell Biology and Immunology, University of StuttgartInstitute of Cell Biology and Immunology, University of StuttgartInstitute of Cell Biology and Immunology, University of StuttgartAbstract Fusion proteins combining TNF-related apoptosis inducing ligand (TRAIL) and antibody building blocks have emerged as a strategy for the targeted treatment of cancer cells. Using a single-chain derivative of homotrimeric TRAIL (scTRAIL), several targeted and non-targeted scTRAIL fusion proteins of varying geometries and valencies for TRAIL receptors and target antigens, all comprising an Fc region, were generated. These fusion proteins comprised either 1 or 2 scTRAIL units, i.e. are tri- or hexavalent for TRAIL receptors and in the targeted versions, 1 or 2 binding sites for EGFR. These fusion proteins were analyzed for cell binding and cell death induction using the EGFR-expressing colorectal cancer cell lines Colo205 and HCT116. In line with previous findings, all fusion proteins that were hexavalent for TRAIL receptors exhibited a strongly increased cell killing activity compared to the trivalent ones. Interestingly, the fusion proteins comprising one scTRAIL unit, did not benefit from targeting to EGFR. In contrast, the hexavalent scTRAIL fusion proteins further benefited from EGFR targeting, resulting in an approximately 6- to 30-fold increase in cell killing. In summary, this study shed further light on the influence of geometry and valency of TRAIL fusion proteins and confirmed IgG-scTRAIL fusion proteins as highly potent cell death inducers.https://doi.org/10.1038/s41598-025-00476-7TNF-related apoptosis-inducing factorTRAILScTRAILAntibody fusion proteinTargetingCancer
spellingShingle Dennis Michler
Oliver Seifert
Klaus Pfizenmaier
Roland E. Kontermann
Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors
Scientific Reports
TNF-related apoptosis-inducing factor
TRAIL
ScTRAIL
Antibody fusion protein
Targeting
Cancer
title Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors
title_full Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors
title_fullStr Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors
title_full_unstemmed Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors
title_short Comparison of antibody-scTRAIL Fc fusion proteins with varying valency for EGFR and TRAIL receptors
title_sort comparison of antibody sctrail fc fusion proteins with varying valency for egfr and trail receptors
topic TNF-related apoptosis-inducing factor
TRAIL
ScTRAIL
Antibody fusion protein
Targeting
Cancer
url https://doi.org/10.1038/s41598-025-00476-7
work_keys_str_mv AT dennismichler comparisonofantibodysctrailfcfusionproteinswithvaryingvalencyforegfrandtrailreceptors
AT oliverseifert comparisonofantibodysctrailfcfusionproteinswithvaryingvalencyforegfrandtrailreceptors
AT klauspfizenmaier comparisonofantibodysctrailfcfusionproteinswithvaryingvalencyforegfrandtrailreceptors
AT rolandekontermann comparisonofantibodysctrailfcfusionproteinswithvaryingvalencyforegfrandtrailreceptors