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Analysis Affinity of Anti-IL-1b Antibody (XMA005/XOMA052) by KinExA (CAT#: STEM-MB-0013-CJ)

Introduction

Interleukin-1β (IL-1β) is a potent multipotent cytokine that affects the function of almost all cell types. IL-1β is a major mediator of inflammatory responses by inducing the growth and differentiation of immunoactive lymphocytes. In this function, IL-1β plays a central role in the prevention of microbial pathogens and tissue damage repair. The expression level and function of IL-1β are strictly regulated by the complex system of IL-1 family members and their receptors. Overexpression of IL-1β is the main cause of increased erythrocyte sedimentation rate (ESR) , peripheral neutrophilia, thrombocytosis, pain and allergy, and anemia. These diseases include systemic juvenile idiopathic arthritis (sJIA) , neonatal multisystemic inflammatory disease (NOMID) , Muckle-Wells syndrome (MWS) , suppurative arthritis, pyoderma gangrena and acne syndrome (PAPA syndrome) , familial Mediterranean fever (FMF) , and others. Excess IL-1β also causes bone degeneration in the joints of patients with rheumatoid arthritis (RA) and affects beta cells in the pancreas, disrupting insulin production in models of type 2 diabetes. Blocking IL-1β has been shown to improve blood glucose control and beta cell function in clinical trials in patients with type 2 diabetes.

XOMA052 is a high affinity, IL-1β-specific therapeutic antibody, also known as gevokizumab. Designed specifically for high potency and infrequent dosing, the antibody is generated from a synthetic mouse antibody sequence and constructed through rational design using XOMA's antibody technology platform. XMA005 is the parent of XOMA052.




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