Bioinspired fabrication of bifunctional antibody–enzyme co-assembled nanocomposites for chemiluminescence immunoassays of E. coli O157:H7

Abstract

Immunoassays based on enzyme-labeled antibodies have been widely used in the food safety field. However, the production process of enzyme-labeled antibodies is complicated and the low storage stability limits their application. Herein, antibody–horseradish peroxidase (HRP) co-assembled nanocomposites (AHC NCs) with outstanding advantages such as enhanced stability, lower cost, and substrate affinity were successfully prepared via a one-pot green method. Then the AHC NCs were employed as an alternative to traditional enzyme-labeled antibodies to develop a chemiluminescence enzyme immunoassay (CLEIA) toward Escherichia coli (E. coli) O157:H7. Under optimal conditions, E. coli O157:H7 can be detected in a linear range from 1 × 10³ CFU mL⁻¹ to 5 × 10⁶ CFU mL⁻¹, while the limit of detection (LOD) is as low as 2.2 × 10² CFU mL⁻¹ (3σ). A series of repeatability studies showed reproducible results with a coefficient of variation of less than 7%. In addition, the proposed CLEIA was successfully applied to the analysis of spiked samples (tap water) and gave quantitative recoveries from 93.72% to 100.72%. This work demonstrates that the developed CLEIA can be applied as a universal platform for specific detection of diversified analytes.