Ranibizumab (Lucentis) is a recombinant human IgG1 monoclonal antibody fragment (Fab) that blocks angiogenesis by inhibiting vascular endothelial growth factor-A (VEGF-A) isoforms. The humanized anti-VEGF monoclonal antibody, Ranibizumab, has been approved by the FDA for treatment of patients with wet age-related macular degeneration. Age-related macular degeneration (AMD) is the leading cause ofirreversible blindness in people over the age of 50 in the developedworld [1]. Currently, the most commonlyused VEGF antagonists are ranibizumab (Lucentis, Genentech, Inc., South San Francisco, CA) and bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA). Ranibizumab, which is an antibody fragment form the bevacizumab molecule with an increased binding affinity for all forms of VEGF, has been approved for the treatment of patients with neo-vascular AMD by the Food and Drug Administration and by the European Medicines Agency since 2006 and 2007, respectively. The DeQuanto Ranibizumab (Lucentis) PK ELISA kit is an ELISA based immunoassay. Standards and diluted samples (serum/ plasma) are incubated in the polystyrene microtiter plate pre coated with recombinant Human vascular endothelial growth factor-A (rhVEGF-A). The Ranibizumab present in the samples binds to the coated rhVEGF-A. Nonbound or the excess substances are removed by washing. Horseradish Peroxidase (HRP) labeled antibody is added to the wells which binds to the rhVEGF-A-Ranibizumabcomplex 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 Ranibizumab 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 Ranibizumab 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 4.88 ng/ml MRD: 1/200

DILUTION LINEARITY: Tested and passes with %RE 20%

ASSAY RANGE: 625 ng/ml to 2.44 ng/ml

ASSAY TIME: 3-4 Hours