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A differential cell capture assay for evaluating antibody interactions with cell surface targets.
David J Sherman1, Vania E Kenanova, Eric J Lepin
1Department of Molecular and Medical Pharmacology, Crump Institute of Molecular Imaging, California NanoSystems Institute, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
A novel cell capture assay allows rapid, low-cost evaluation of antibody binding to cell surface targets. This benchtop method correlates antibody concentration-dependent cell capture with binding affinity, aiding antibody selection for targeting studies.
Area of Science:
- Biotechnology
- Immunology
- Cell Biology
Background:
- Conventional binding assays like ELISA and flow cytometry present challenges including high cost, labor intensity, and significant reagent/sample consumption.
- There is a need for accessible, benchtop tools to quickly assess antibody binding efficacy to cell surface targets.
- Antibodies and antibody fragments are crucial reagents in biological and biomedical laboratory assays for targeting cell surface molecules.
Purpose of the Study:
- To develop and validate a novel, cost-effective cell capture assay for evaluating the relative binding capabilities of antibodies and antibody fragments to cell surface targets.
- To demonstrate the assay's utility in comparing the binding affinities of different antibodies against a specific cell surface target.
- To provide a benchtop alternative to existing complex and expensive binding assays.
Main Methods:
- A novel cell capture assay was developed utilizing a functionalized microscope slide surface.
- Chimeric rituximab and murine anti-CD20 monoclonal antibodies were employed as capture agents to bind CD20-expressing mammalian cells.
- The relative binding affinities of antibodies were assessed by quantifying the concentration-dependent cell capture profiles.
Main Results:
- The cell capture assay successfully enabled rapid evaluation of relative antibody binding efficacies.
- Antibody concentration-dependent cell capture profiles were found to correlate directly with their relative binding affinities.
- The impact of different capture surfaces on assay outcomes was identified as a key factor.
Conclusions:
- The developed cell capture assay offers a quick, low-cost, benchtop method for assessing antibody binding to cell surface targets.
- This assay provides a valuable tool for selecting antibodies or antibody fragments for specific cell targeting applications.
- The findings highlight the assay's potential to streamline antibody screening processes in biological and biomedical research.
