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Analysis Affinity of TAK-079 (Human) by KinExA (CAT#: STEM-MB-0010-CJ)

Introduction

TAK-079 is a fully human, non-excitatory, IgG1 cell-depletion monoclonal antibody that binds to human CD38 with high affinity and cross-reacts with CD38 in cytophagus monkeys. TAK-079 binding to CD38 does not result in PBMC cytokine activation, but it mediates the depletion of human B-line cells by ADCC and CDC, and in most cases, lines with increased CD38 expression are more susceptible to cell lysis. In vivo, TAK-079 effectively inhibited the antigen memory response of human B cells in a mouse adoptive transfer model.




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

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: Serum, Plasma, Urine
• Equipment: Kinetic Exclusion Assay (KinExA)