Analysis Ligand-receptor Binding of Biopharmaceutical Compounds by Kinetic Exclusion Assay (CAT#: STEM-B-0392-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

KinExA is a two-stage analysis system. In the first stage, a number of solutions are prepared, where one partner remains constant (constant binding partner, or CBP) and the other (titrant) is variable, usually serially diluted. As the titrant is added, the free CBP decreases and is analysed by a sophisticated and precise microfluorescence measurement device. The signal generated can be mathematically related to the affinity (KD) of the two molecules for each other, as well as the kinetic parameters of binding (kon) and dissociation (koff).

Applications

Biopharmaceutica

Procedure

1. preparation of the functionalized beads which will capture the analyte for measuremen.
2. preparation of a series of solutions consisting of a constant initial concentration of one component of the binary reaction and serial dilutions of the other reactant. The component that is kept constant is the constant binding partner (CBP) , and is the one which will be analyzed.
3. each reaction mixture is sampled and the fluorescence of free CBP bound to the capture beads is obtained for subsequent numerical analysis.

Materials

• Sample: Peptides, Proteins, Vaccines, Virus-like particles
• Equipment: Kinetic Exclusion Assay (KinExA)

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