Peptide Functionalized Gold Nanoparticles with Optimized Particle Size and Concentration for Colorimetric Assay Development: Detection of Cardiac Troponin I

Peptide-functionalized gold nanoparticles (AuNPs) are extensively utilized in colorimetric assays for rapid and sensitive detection of various biomedical and environmental targets. Although extensively used as colorimetric reporting systems, the role of the size and concentration of the AuNPs has not been thoroughly investigated. In this study, a 12-mer cardiac troponin I (cTnI)-specific peptide CALNN-Peg₄-FYSHSFHENWPS was immobilized on AuNPs of different size and concentration via the CALNN anchoring sequence. A relationship was established between the total surface area of the AuNPs (binding availability) and response (centroid shift). Moreover, a colorimetric assay for cTnI operating under optimized conditions (36 nm AuNPs) yielded a limit of detection of 0.2 ng/mL (8.4 pM) when tested in diluted serum samples with an assay time of 10 min. This encouraging result opens up for further development of AuNP assays in early diagnosis of cardiac injury.