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A Flow Cytometry-Based Cell Surface Protein Binding Assay for Assessing Selectivity and Specificity of an Anticancer Aptamer
Published on: September 13, 2022
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Selection and characterisation of Affimers specific for CEA recognition
Shazana Hilda Shamsuddin1,2, David G Jayne3, Darren C Tomlinson4,5
1The Leeds Bionanotechnology Group, School of Biomedical Sciences, University of Leeds, Leeds, LS2 9JT, UK. shazana.hilda@usm.my.
Scientific Reports
|January 13, 2021
Summary
Researchers developed novel Affimer proteins for detecting carcinoembryonic antigen (CEA), a key biomarker for colorectal cancer (CRC). These Affimers offer a promising, antibody-free alternative for sensitive and specific CRC diagnostics and therapies.
Area of Science:
- Biotechnology
- Biochemistry
- Molecular Biology
Background:
- Carcinoembryonic antigen (CEA) is a crucial blood biomarker for colorectal cancer (CRC) screening and recurrence monitoring.
- Current diagnostic methods for CRC face limitations in sensitivity, specificity, and cost-effectiveness.
- Existing antibody-based detection systems have drawbacks, including multimeric binding, high cost, and production challenges.
Purpose of the Study:
- To develop Affimer proteins as a novel, antibody-independent binding reagent for CEA detection.
- To address the limitations of current CRC diagnostic tools by creating a more sensitive and specific detection method.
- To explore the potential of Affimers in various applications, including diagnostics, therapy, and biosensing.
Main Methods:
- Affimer proteins specific to CEA were selected from a phage display library.
- The binding specificity and selectivity of anti-CEA Affimers were characterized using cell culture lysates and fixed cells.
- Surface Plasmon Resonance (SPR) was employed to analyze the kinetic binding properties and affinity of the Affimers.
Main Results:
- Three anti-CEA Affimer proteins were successfully selected and characterized.
- The Affimers demonstrated specific and selective binding to CEA protein epitopes.
- Affimers exhibited high-affinity binding to CEA in the nanomolar (nM) range, confirmed by SPR analysis.
Conclusions:
- Affimer proteins represent a viable and advantageous alternative to antibodies for CEA detection.
- These novel Affimers possess significant potential for use in advanced colorectal cancer diagnostics and targeted therapies.
- The developed Affimers are suitable for applications in diagnostic imaging, affinity purification, and biosensor development.
