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Updated: Jan 8, 2026
A Flow Cytometry-Based Cell Surface Protein Binding Assay for Assessing Selectivity and Specificity of an Anticancer Aptamer
Published on: September 13, 2022
Preclinical Proof of Concept for the Single-Protein Anticancer Molecule Targeting Both a Tumor Surface Antigen and an
Kyu Tae Byun1, Boram Kim1, Inbeom Lee1
1Department of Biotechnology and Research Institute for Biomedical and Health Science, College of Biomedical and Health Science, Konkuk University, Chungju, Republic of Korea.
Abstract:
Although antibody-drug conjugates (ADCs) are a widely used platform for developing various anticancer immunotherapeutics, the use of cancer non-selective chemicals is recognized as a drawback of ADC development. To address the issues regarding the safety and manufacturing complexity of ADCs, this study conceptualizes a single-protein platform, named dual-targeting anticancer therapeutics (DTAT), that links a cancer cell-selective cytotoxic peptide to an antibody via a linker peptide cleavable on cancer cells. As a model molecule for preclinical proof of the concept, an anti-Her2 single-chain variable fragment (scFv)-based DTAT protein named DTAT-D311 is established, which contains a recently developed anticancer peptide, herein named CPTin, as its cell-penetrating payload. This recombinant single protein efficiently induces the apoptotic death of cancer cells, which is characterized by a very early onset. In terms of in vivo efficacy in suppressing tumor growth, DTAT-D311 outperforms the anti-Her2 therapeutic antibody, trastuzumab (Herceptin). By targeting an intracellularly addictive oncoprotein, CP2c, CPTin exhibits broad-spectrum anticancer activity. In conclusion, this study demonstrates that DTAT provides an innovative pharmaceutical modality to target both a tumor surface antigen and an intracellular oncoprotein. In addition, DTAT-D311 is suggested to be a promising biopharmaceutical agent for targeted immunotherapy against Her2-positive cancers, exhibiting favorable safety profile.
Insights
This study introduces dual-targeting anticancer therapeutics (DTAT), a novel single-protein platform that improves upon antibody-drug conjugates (ADCs) by enhancing safety and simplifying manufacturing for targeted cancer immunotherapy.
Area of Science:
- Biotechnology
- Immunotherapy
- Oncology
Background:
- Antibody-drug conjugates (ADCs) face challenges with non-selective chemical use and manufacturing complexity.
- Developing safer and more effective targeted cancer immunotherapeutics is a critical need.
Purpose of the Study:
- To develop a novel single-protein platform, dual-targeting anticancer therapeutics (DTAT), addressing ADC limitations.
- To establish and evaluate DTAT-D311, an anti-Her2 scFv-based DTAT, for preclinical proof of concept.
Main Methods:
- Conceptualized and engineered a DTAT protein linking a cytotoxic peptide to an antibody via a cleavable linker.
- Utilized an anti-Her2 single-chain variable fragment (scFv) and a novel anticancer peptide (CPTin) as payload.
- Assessed in vitro cancer cell apoptosis induction and in vivo tumor growth suppression.
Main Results:
- DTAT-D311 demonstrated efficient and early-onset apoptotic cancer cell death.
- DTAT-D311 showed superior in vivo tumor growth suppression compared to trastuzumab.
- CPTin exhibited broad-spectrum anticancer activity by targeting the intracellular oncoprotein CP2c.
Conclusions:
- DTAT platform offers an innovative approach targeting both surface antigens and intracellular oncoproteins.
- DTAT-D311 is a promising biopharmaceutical for Her2-positive cancers, with a favorable safety profile.
- This modality enhances targeted immunotherapy for improved cancer treatment.
