Denosumab is a human monoclonal antibody used for treatment of osteoporosis. Denosumab binds to RANKL, a transmembrane protein which is essential for formation, function and survival of osteoclasts. Drug level quantification can be important to adapt patient prescription or to switch to an alternative drug. The DeQuanto Denosumab PK ELISA kit is an ELISA based immunoassay. The Denosumab present in the sample binds to coated Anti Denosumab antibody on polystyrene microtiter plate. Non bound or the excess substances are removed by washing. Horseradish Peroxidase (HRP) labeled antibody is added to the wells which binds to the complex formed in the wells. Following a wash to remove any unbound antibody-enzyme reagent, substrate solution is added to the wells. A color product is formed in proportion to the amount of denosumab present in the sample or standards. The color development is stopped by addition of a stop solution. The absorbance is measured at 450 nm in a microtiter plate reader. The concentration of denosumab in a sample can be interpolated from the standard curve.

ACCURACY AND PRECISION: Inter and intra assay precision %RE 20%

SELECTIVITY: Tested in Human Plasma Matrices

SENSITIVITY: LLOQ is 3.9 ng/ml MRD: 1/200

DILUTION LINEARITY: Tested and passes with %RE 20%

ASSAY RANGE: 500 ng/ml to 3.9 ng/ml

ASSAY TIME: 3-4 Hours