Analysis Ligand-receptor Binding of Biopharmaceutical Compounds by Fluorescence Resonance Energy Transfer (FRET) (CAT#: STEM-B-0394-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.

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Principle Applications Procedure Materials Notes Related Products

Principle

Fluorescence Resonance Energy Transfer (FRET) utilizes energy transferred between the donor and the acceptor molecules that are in close proximity. FRET uses a fluorescently labeled ligand, as with FP. Energy transfer within FRET begins by exciting the donor. The dipole-dipole interaction between the donor and the acceptor molecule transfers the energy from the donor to the acceptor molecule. If the ligand is bound to the receptor-antibody complex, then the acceptor will emit light.

Applications

Biopharmaceutica

Procedure

1. Prepare samples.
2. Put the sample into the equipment and run the instrument according to the instruction manual.
3. Collect and analyze data.

Materials

• Sample: Peptides, Proteins, Vaccines, Virus-like particles
• Equipment: Fluorescence Resonance Energy Transfer (FRET) Microscopy

Notes

When using FRET, it is critical that there is a distance smaller than 10 nm between the acceptor and donor, in addition to an overlapping absorption spectrum between acceptor and donor, and that the antibody does not interfere or block the ligand binding site.

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