Analysis Affinity of Human Monoclonal Antibody to CD22 by KinExA (CAT#: STEM-MB-0031-CJ)

Introduction

CD22 is a 135-kDa transmembrane sialoglycoprotein and a member of the immunoglobulin superfamily. Its expression is restricted to lymphocytes of the B cell lineage and is highly developmentally regulated: CD22 is present in the cytoplasm of pro- and pre-B cells and becomes detectable on the cell surface only at mature stages of B cell differentiation. Cell surface expression is lost during terminal differentiation into plasma cell and after B cell activation.CD22 is also expressed by the vast majority of B cell NHLs.

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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

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, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)

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