In a new study, researchers from King Saud University have developed a novel ELISA assay with high sensitivity and selectivity for bioanalysis of bevacizumab (BEV), a monoclonal antibody used for immunotherapy of different types of cancer. The work was published in the Current Analytical Chemistry.
The group designed the ELISA to capture BEV by using human vascular endothelial growth factor protein (VEGF) that is immobilized onto a 96-well ELISA plate. The BEV-VEGF complex is formed onto the plate wells and is then quantified using horseradish peroxidase labeled anti-human IgG (HRP-IgG) and 3,3`,5,5`-tetramethylbenzidine (TMB) as a chromogenic substrate for peroxidase enzyme.
The assay has been validated for its accuracy and precision. The authors report that the assay is easy and can analyze a batch of ~ 200 samples, making it ideal for a clinical setting. In addition, with this assay, the pretreatment of plasma samples by affinity chromatography or other sophisticated equipment is no longer required and can estimate the nano-gram level of BEV.
The ELISA for BEV is expected to contribute in studying its PK, PD, TDM and assessing the expected bioavalilability of biosimillars or biobetters.
Image: Schematic diagram of the ELISA for Bevacizumab. Image courtesy of Manal A. El-Gendy et al.