Abstract
The enzyme-linked immunosorbent assay (ELISA) remains the prevailing method for quantifying protein biomarkers. Enzymatic signal generation and amplification are key mechanisms that govern its analytical performance. This study reports the synthesis and application of microscale metal-organic framework (MOF)/enzyme composite particles as a novel detection probe to substantially enhance the sensitivity of ELISA. An optimal one-pot approach was established to incorporate a substantial amount of streptavidin-horseradish peroxidase (SA-HRP) either within or on the surface of the metal-azolate framework (MAF-7) microparticles. This approach enables the labeling of a single sandwich antibody-antigen complex with numerous enzymes, which markedly amplifies the enzymatic colorimetric signal generation. Moreover, MAF-7 caging was found to enhance the reactivity of the caged HRP enzyme, further promoting the overall detection sensitivity of ELISA. Compared to other developments that are often associated with more complicated detection modalities, our method is compatible with standard immunoassays and commonly used photometrical signal detection. The implementation of this strategy in the detection of CD147 results in a remarkably low limit of detection of 2.8 fg mL(-1), representing a 10(5)-fold improvement compared to that obtained with the standard ELISA. Moreover, the heightened sensitivity of this technique renders it particularly suitable for diagnosing breast cancer, thus presenting a promising tool for the early detection of the disease in clinical settings.