A High-Throughput Platform for the Rapid Quantification of Phosphorylated Histone H2AX in Cell Lysates Based on Microplate Electrochemiluminescence Immunosensor Array

Sensitive detection of phosphorylated histone H2AX (γH2AX) in cells as a biomarker of DNA double-strand breaks has great significance in the field of molecular toxicology and life science research. However, current γH2AX detection methods require labor- and time-consuming steps. Here, for the first time, we designed a simple electrochemiluminescence (ECL) immunoassay integrated with a microplate-based sensor array to realize sensitive and high-throughput detection of γH2AX in cell lysates. Under the optimized conditions, this ECL immunosensor array could linearly respond to γH2AX concentrations in the range from 2 × 10² to 1 × 10⁵ pg/mL. In addition, our approach possessed excellent specificity and satisfactory reproducibility, and its practicality was verified in real cell lysates. The whole process including instrumental and manual operation was completed in no more than 3 h. This study provides a convenient and rapid alternative method for the sensitive quantification of γH2AX, which shows promising application in high-throughput screening of genotoxic chemicals and drug candidates.