Desarrollo de nanobiomarcadores fluorescentes de puntos cuánticos de carbono-IgG humana para inmunodiagnósticos...

Rapid and sensitive detection of pathogen-specific antigens or antibodies through point-of-care testing (POCT) is crucial for effective infectious disease diagnosis and surveillance. This study reports the development of a robust fluorescent nanobioprobe utilizing green-synthesized heteroatom (N, K, Ca) doped carbon quantum dots (CQDs) covalently conjugated with human/goat IgG antibodies. Amine- and carboxyl-functionalized CQDs were hydrothermally synthesized from Cissus quadrangularis (cq) extract and comprehensively characterized by HR-TEM, XPS, FTIR, NMR, UV-Vis, and fluorescence spectroscopy. Amine-functionalized CQDs exhibited ∼1.9-fold enhanced emission fluorescence at 466 nm wavelength due to nitrogen-induced charge stabilization and abundant surface functional groups. Unlike conventional carboxyl-QD/amine-IgG systems, this study demonstrated amine-CQDs/carboxyl-IgG nanobioprobes with optimal fluorescence at 0.2 mg/mL IgG concentration. HRTEM, FTIR, and zeta potential post conjugation confirmed stable IgG conjugation. Fluorescence response of cq-hCQDs-IgG(G/H) nanobioprobes demonstrated a concentration-dependent (0.01-0.1 mg/mL) increase with emission at 466 nm. IgG-(H) immunoreactivity post-conjugation was further validated using FMA and ELISA showing a linear increase in fluorescence response for binding. Limit of detection (LOD) 2.65 ng and limit of quantification (LOQ) 8.85 ng was estimated for F'_ab specific anti-human IgG. Sensitivity of the nanobioprobe was evaluated by spot assay on a nitrocellulose membrane under UVA excitation (365 nm). Smartphone-enabled analysis yielded a linear detection range of 0.01-0.1 mg/mL with LOD/LOQ values of 22.7/68.8 μg/mL (human) and 28.5/86.4 μg/mL (goat). In addition, cq-hCQDs-IgG(H) demonstrated strong photostability with only a 2.14% G-value loss after 60 days, while greenness assessment confirmed the environmentally sustainable nature of both the CQDs and their bioconjugates, thereby underscoring the potential of cq-hCQDs-IgG nanobioprobes as promising probes for immunodiagnostics.