Prolia RANK IκB Neutralisation Assay

For the evaluation of the biological activity of denosumab innovator and biosimilar materials, BioOutsource offers an off-the-shelf reporter gene bioassay (DiscoverX).  Denosumab is a fully human sequence derived monoclonal antibody utilized in the treatment of osteoporosis via the inhibition of the RANK (Receptor Activator of Nuclear Factor-kappa B) pathway.  In this assay design, a transgenic U2OS cell line expresses RANK and an IκB-enzyme donor fusion protein.  Upon exposure to the enzyme acceptor in the substrate, the active enzyme is formed which hydrolyses the substrate and produces a chemiluminescent signal. RANKL binds to RANK receptor on the cell surface resulting in NF-κB signalling and IκB degradation, leading to a decrease in chemiluminescent signal.  Denosumab inhibits RANKL-based activation of RANK, leading to an increase in chemiluminescence. The increase in chemiluminescence is directly proportional to the functional activity of denosumab.

 

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