Unlock Exclusive Discounts & Flash Sales! Click Here to Join the Deals on Every Wednesday!

Analysis Affinity of IL-1β ± XOMA 052 to Soluble IL-1 Receptors sRI and sRII by KinExA (CAT#: STEM-MB-0042-CJ)

Introduction

IL-1β is a highly potent cytokine that drives the acute phase inflammatory response and has an essential role in the innate immune response. As in many receptor-ligand systems, IL-1β signaling is complex, with multiple ligands interacting with membrane-bound and soluble forms of several receptors. The complexity of receptor-ligand system presents a challenge for the selection of therapeutic anti-IL-1β antibodies. The optimal antibody would selectively attenuate systemic high-level IL-1β signaling to lower, beneficial levels while allowing very high local concentrations of IL-1β to initiate protective inflammatory responses (in response to infection, for instance).<br /><br />XOMA 052, a recombinant monoclonal antibody with long circulating half-life, high affinity, and high specificity for IL-1β, that simultaneously reduces the affinity of IL-1β for IL-1RI (K′RL >KRL) while leaving affinity for IL-1RII and the soluble inhibitory receptors largely unaffected by antibody binding. XOMA 052 is a potent inhibitor of IL-1β activity both in vitro and in vivo.




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: Antibodies, Cells, Small Molecules, Proteins, DNA, Lipids, Viruses, Serum, & More
• Equipment: Kinetic Exclusion Assay (KinExA)