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Analysis Affinity of Protein to Recombinant Cynomolgus P-cadherin (human,monkey) by KinExA (CAT#: STEM-MB-0043-CJ)

Introduction

Classical cadherins (E-, N-, and P-cadherin) comprise a family of molecules that mediate calcium-dependent cell-cell adhesion and are localized at the adherens junctions. P-cadherin overexpression has correlate with increased tumor cell motility and invasiveness. Upregulation of P-cadherin has been reported in various tumors, including breast, gastric, endometrial, colorectal and pancreatic cancers, and is correlated with poor survival of breast cancer patients. In contrast, low levels of the P-cadherin gene expression have been detected in normal tissues. P-cadherin therefore represents an interesting target for the treatment of solid tumors that show increased levels of expression.Bispecific antibodies (bsAb) have emerged in recent years as promising agents for immune-mediated tumor cell killing. The approval by the US E64of the bispecific T cell engager blinatumomab for the treatment of relapsed or refractory B cell precursor acute lymphoblastic leukemia.




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; 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)
• Optionals: CHO Cells
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