Multilayered Mesoporous Composite Nanostructures for Highly Sensitive Label-Free Quantification of Cardiac Troponin-I

Cardiac troponin-I (cTnI) is a well-known biomarker for the diagnosis and control of acute myocardial infarction in clinical practice. To improve the accuracy and reliability of cTnI electrochemical immunosensors, we propose a multilayer nanostructure consisting of Fe₃O₄-COOH labeled anti-cTnI monoclonal antibody (Fe₃O₄-COOH-Ab₁) and anti-cTnI polyclonal antibody (Ab₂) conjugated on Au-Ag nanoparticles (NPs) decorated on a metal-organic framework (Au-Ag@ZIF-67-Ab₂). In this design, Fe₃O₄-COOH was used for separation of cTnI in specimens and signal amplification, hierarchical porous ZIF-67 extremely enhanced the specific surface area, and Au-Ag NPs synergically promoted the conductivity and sensitivity. They were additionally employed as an immobilization platform to enhance antibody loading. Electron microscopy images indicated that Ag-Au NPs with an average diameter of 1.9 ± 0.5 nm were uniformly decorated on plate-like ZIF-67 particles (with average size of 690 nm) without any agglomeration. Several electrochemical assays were implemented to precisely evaluate the immunosensor performance. The square wave voltammetry technique exhibited the best performance with a sensitivity of 0.98 mA mL cm^-2 ng^-1 and a detection limit of 0.047 pg mL^-1 in the linear range of 0.04 to 8 ng mL^-1.