Bevacizumab (Avastin) is a recombinant human IgG1 monoclonal antibody specific for all human vascular endothelial growth factor (VEGF) isoforms. The humanized anti-VEGF monoclonal antibody, bevacizumab, has been approved by the FDA as a first-line treatment for metastatic colorectal cancer in combination with chemotherapy. The pharmacokinetic properties of bevacizumab in several species have been previously described and are consistent with a typical humanized monoclonal antibody. It was shown in the literature that the surveillance of circulating concentration during maintenance therapy represents a direct and/or indirect factor for some other side effects. Identification of biomarkers for (non-)response and risk factors for adverse drug reactions that might be related to serum concentrations and maintaining the effective concentration of Bevacizumab in order to potentially avoid some side effects with a reliable method might be beneficial. The DeQuanto Bevacizumab (Avastin) 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 (rhVEGF). The Bevacizumab present in the samples binds to the coated rhVEGF. Non bound or the excess substances are removed by washing. Horseradish Peroxidase (HRP) labeled antibody is added to the wells which binds to the rhVEGF-Bevacizumab complex 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 Bevacizumab 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 Bevacizumab 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 1.17 ng/ml MRD: 1/300

DILUTION LINEARITY: Tested and passes with %RE 20%

ASSAY RANGE: 1200 ng/ml to 1.17 ng/ml

ASSAY TIME: 3-4 Hours