Antigen capture ELISA for PK assay developmentEnlarge
A microtiter plate was coated overnight with VEGF at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, 10% cynomolgus macaque serum was added spiked with increasing concentrations of bevacizumab (hIgG1/kappa). Detection was performed using either HRP conjugated Human Anti-Human hIgG1 (Fc) (clone AbD27686, HCA285P), HRP conjugated Mouse Anti-Human IgG (CH2 Domain) Antibody (clone R10Z8E9, MCA5748G) or, as a control for an antibody that is not human Fc specific, Mouse Anti-Human IgG (Fc) CH2 Domain:HRP (clone MK 1 A6, MCA647P) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.