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Analysis Affinity of Human mAb GNCgzk to Cocaine by KinExA (CAT#: STEM-MB-0032-CJ)

Introduction

Cocaine functions as a molecular blocker of dopamine transporters, and thus drug detoxification within the CNS via its displacement from transporter binding sites poses an interesting conundrum. Herein, active and passive vaccination strategies that yield therapeutic levels of cocaine-binding antibodies have been explored in rodent models for drug detoxification as well as for the prevention of drug acquisition, self-administration, and reinstatement.GNCgzk is a monoclonal antibody against cocaine. Preclinical validation of GNCgzk for cocaine-related indications.




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

Immunology/Inflammation, Toxicology; 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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