Preparation of an Acridinium Ester-Labeled Antibody and Its Application in GoldMag Nanoparticle-Based, Ultrasensitive Chemiluminescence Immunoassay for the Detection of Human Epididymis Protein 4

An ultrasensitive and rapid sandwich-type chemiluminescence immunoassay (CLIA) was developed for the clinical determination of human epididymis protein 4 (HE4) in human serum, using GoldMag nanoparticles as solid phase and acridinium ester (AE) as chemiluminescence system (GMP-CLIA). The process of AE labeling antibodies was systematically studied and evaluated. The effect of varies factors such as molar ratio of AE to antibodies, labeling time, and the components of elution buffer and trigger solution were optimized. Under the selected conditions, AE labeling experiments were successfully performed with the average labeling efficiency of 1.92 ± 0.08, and antibody utilization rate of 69.77 ± 1.19%. Antibody activity remained unchanged after labeling. The established GMP-CLIA method can detect HE4 in the range of 0.25–50 ng·mL−1 (10–2000 pM) with a detection limit of 0.084 ng·mL−1 (3.36 pM). The sensitivity has reached a high level, comparable with the current commercial detection kits. This proposed method has been successfully applied to the clinical determination of HE4 in 65 human sera. The results showed a good correlation with a clinical method, microplate-based chemiluminescence enzyme immunoassay (CLEIA), with the correlation coefficient of 0.9594.