Our expert team will engage in a detailed discussion to understand your specific research objectives, target characteristics, and therapeutic candidates. Based on this, we will design a customized cell-based binding assay strategy tailored to your project's unique requirements, selecting the most appropriate assay format and detection method.
Cell based Binding Assay
Overview
The landscape of therapeutic development, particularly for biologics, demands assays that accurately reflect in vivo interactions. Traditional binding assays, often relying on purified recombinant proteins, can overlook critical factors such as native protein conformation, post-translational modifications, and the complex cellular environment. This can lead to misleading data and delays in drug development. Cell-based binding assays are essential because they provide a physiologically relevant platform to assess therapeutic binding to targets in their native state on the cell surface, crucial for understanding functional activity and ensuring the translatability of preclinical findings to clinical outcomes.
Fig.1 Assay for RBD binding avidity.1,3
Cell based Binding Assay at Creative Biolabs
Creative Biolabs' cell-based binding assay service provides precise, physiologically relevant data critical for advancing your therapeutic candidates. We offer comprehensive solutions for characterizing antibody-target interactions, assessing functional binding, and streamlining your screening processes. Our deliverables include robust binding profiles, detailed kinetic data, and insights into the specificity and affinity of your molecules, all grounded in a native cellular context.
Required Starting Materials
To initiate your project, we typically need our customer to provide the following:
- Target Protein Information: Detailed sequence information and any known expression systems for your target.
- Antibody/Ligand Candidates: Purified antibodies, crude samples, or hybridoma supernatants for analysis.
- Specific Cell Lines: If you have proprietary cell lines expressing your target, otherwise, we can utilize our extensive internal library.
Our Service Workflow: From Concept to Clinical
Project Consultation & Assay Design
Cell Line Engineering/Selection
We leverage our extensive repository of over 100 high-standard cell lines. If your target is not expressed in our existing lines, our advanced cell engineering capabilities allow us to develop custom stable cell lines expressing your specific target protein on the cell surface, ensuring optimal assay performance.
Assay Optimization & Validation
Before full-scale execution, we meticulously optimize all critical assay parameters, including cell density, antibody/ligand concentration, incubation times, and buffer conditions. This step is followed by rigorous validation to ensure the assay's sensitivity, specificity, reproducibility, and robustness, guaranteeing high-quality data.
Binding Assay Execution
Our skilled scientists then execute the chosen cell-based binding assay (e.g., Flow Cytometry, ELISA, or competitive assays) on your provided samples using state-of-the-art instrumentation and strict quality control measures. Each experiment is performed with precision to generate reliable raw data.
Data Analysis & Reporting
Following assay execution, comprehensive data processing and interpretation are performed. We generate detailed reports that include all experimental parameters, raw data, processed data tables, and expertly interpreted binding curves (e.g., EC50, KD values). Our reports provide clear conclusions and actionable insights for your project.
Assay Approaches List
| Approaches | Readout | |
|---|---|---|
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With the use of a fluorescent secondary antibody, the test sample's capacity (antibody, ADC, etc.) to bind to the target protein on the cell surface can be determined by measuring the specific fluorescence value. | Fluorescence |
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The blocking test involves adding more receptor ligands to the antibody sample so that they can bind with the FACS sample. The specific fluorescence value may be determined using the fluorescent secondary antibody to indicate the blocking activity of the test materials. | |
|
To bond with the target protein, an additional Target Ab will be introduced during the competition test. When antibody samples impede Target Ab's binding to the target antigen, it indicates that the antibodies identify the same epitope. | |
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A direct immunochemical labeling procedure that incorporates cell culture and ELISA. | Absorbance |
Service Highlights
Physiological Relevance
Assess binding in a native cellular environment, providing more accurate in vivo predictions.
No Protein Purification Needed
Eliminates the need for costly and time-consuming recombinant protein expression and purification.
High-Throughput Capability
Efficiently screen large libraries of candidates, accelerating your discovery process.
Functional Insights
Gain a deeper understanding of target engagement and potential functional activity.
Versatility
Compatible with various detection methods and applicable to diverse therapeutic modalities.
Speed
Streamlined workflow and established platforms lead to faster turnaround times.
Published Data
This study introduces a novel, microtiter plate-based CXCR4 antibody competition assay designed to efficiently and affordably determine the inhibitory concentrations (IC50) and affinity constants (Ki) of CXCR4-targeting drugs. The assay leverages the principle that most CXCR4 antagonists compete with a fluorescence-tagged CXCR4 antibody for receptor binding. This method enables rapid measurement of binding affinities for various CXCR4 antagonists directly in living cells, with results obtainable within just three hours.
Fig.2 Cell-based CXCR4 ECL-2 specific antibody competition.2,4
Frequently Asked Questions
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Q1: What types of targets can be analyzed using Creative Biolabs' cell-based binding assays?
A1: We can analyze a wide range of targets, including cell surface receptors, multi-pass membrane proteins, ion channels, and viral proteins expressed on the cell surface. Our assays are particularly valuable for targets that are challenging to express or purify in a functional form.
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Q2: Can Creative Biolabs develop custom cell lines for my specific target if it's not in your existing library?
A2: Absolutely. We specialize in custom cell line development. If your target is not represented in our extensive library of over 100 cell lines, our expert team can engineer stable cell lines expressing your specific protein to ensure optimal assay performance.
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Q3: What is the typical turnaround time for a Cell-Based Binding Assay project?
A3: The timeframe can vary depending on the complexity of your target, the number of samples, and the specific assay requirements. Generally, projects range from 4 to 12 weeks from initial consultation to final report delivery. We will provide a detailed timeline during your project consultation.
If you have any inquiries about our cell-based binding assay and wish to assess your candidates' binding potential to specific targets in a cell-based manner, please do not hesitate to contact us.
References
- Takahashi, Etsuhisa et al. "Clinical Utility of SARS-CoV-2 Antibody Titer Multiplied by Binding Avidity of Receptor-Binding Domain (RBD) in Monitoring Protective Immunity and Clinical Severity." Viruses vol. 15,8 1662. 30 Jul. 2023, doi:10.3390/v15081662.
- Harms, Mirja et al. "Microtiter plate-based antibody-competition assay to determine binding affinities and plasma/blood stability of CXCR4 ligands." Scientific Reports vol. 10,1 16036. 29 Sep. 2020, doi:10.1038/s41598-020-73012-4
- Distributed under Open Access License CC BY 4.0. The original image was modified by extracting and using part A, and the title was changed to "Assay for RBD binding avidity".
- Distributed under Open Access License CC BY 4.0. The original image was modified by extracting and using parts a & b, and the title was changed to "Cell-based CXCR4 ECL-2 specific antibody competition".
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