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Analysis Ligand-receptor Binding of Biopharmaceutical Compounds by Fluorescence Polarization (FP) (CAT#: STEM-B-0391-CJ)

Introduction

Biological activity is, besides the safety of the therapy, the most important aspect of a drug product or therapy. Thus, a major goal of formulation development is to maintain the functionality of the therapeutic compound inside its formulation throughout its shelf-life.

Binding of a ligand to a receptor changes its shape or activity, allowing it to transmit a signal or directly produce a change inside of the cell. Stages of signal transduction: ligand-receptor binding, signal relay, response.




Principle

Fluorescence polarization (FP) technology is based on the measurement of molecule rotation, and has been widely used to study molecular interactions in solution. This method can be used to measure binding and dissociation between two molecules if one of the binding molecules is relatively small and fluorescent.

Applications

Biopharmaceutica

Procedure

1. Estimate the best settings for instrument.
2. Determine the optimal concentration of free tracer.
3. Titrate binder with appropriate controls.
4. Titer competitor in one or more assay formulations.
5. Assay characterization.
6. Troubleshooting.

Materials

• Sample: Peptides, Proteins, Vaccines, Virus-like particles
• Equipment: Fluorescence Polarization (FP)

Notes

This method is less precise at low nanomolar concentrations.
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