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Analysis Affinity and Kinetics of Monoclonal Antibody (mAb) against Membrane Proteins by KinExA (CAT#: STEM-MB-0065-CJ)

Introduction

Monoclonal antibody (mAb) therapeutics have formed and continue to form the vast majority of biopharmaceutical company pipelines today with a number of remarkable commercial successes. The advent of mAbs as therapeutics has been greatly aided by a process platform approach that has enabled rapid development and manufacturing for this class of drugs. Meanwhile monoclonal antibody is an illustration of a remarkable translational scientific discovery and its bench-to-bedside journey took around a decade to be materialized into the clinical application of its use. However, a balance between the cost of treatment, efficacy and safety need to be considered as the treatment with mAbs can be expensive. Hence, more robust data on their cost effectiveness and long-term safety are warranted.




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

Oncology & Cancer; Virology;Pharmacology

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 Type: Antibodies, Cells, Small Molecules, Proteins, DNA, Lipids, Viruses, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)
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