A Novel Highly Sensitive Chemiluminescence Enzyme Immunoassay with Signal Enhancement Using Horseradish Peroxidase-Luminol-Hydrogen Peroxide Reaction for the Quantitation of Monoclonal Antibodies Used for Cancer Immunotherapy

The development and validation of a novel enhanced chemiluminescence enzyme immunoassay (CLEIA) with excellent sensitivity for the quantification monoclonal antibodies (mAbs) used immunotherapy cancer are described in this paper first time. 96-microwell plates were assay procedures, which involved non-competitive binding reaction to specific antigen. immune complex antigen-mAb formed on internal surface plate wells was quantified by (CL)-producing horseradish peroxidase (HRP) reaction. employed 4-(imidazol-1-yl)phenol (IMP) as highly potent signal enhancer HRP-luminol–hydrogen peroxide (H2O2) CL proposed CLEIA developed bevacizumab (BEV), representative example mAbs. validated accordance bioanalysis standards, all criteria met. assay’s limit detection (LOD) quantitation (LOQ) 9.3 28.2 pg mL−1, respectively, working dynamic range 10–400 mL−1. enables accurate precise mAbs human plasma samples without any interference from endogenous substances and/or matrix. compared terms other pre-validated enzyme-linked immunosorbent (ELISA) using HRP/colorimetric substrate system observed differences explained. protocol’s ease use, high throughput, simplicity allows analyze numerous clinical settings. has significant benefit assessment settings evaluation their pharmacokinetics, pharmacodynamics, therapeutic drug monitoring, refining safety profiles, opening new era better understanding pharmacodynamics at cellular level.