Biofabricated functionalized graphene quantum dots (fGQDs): unraveling its fluorescence sensing mechanism of human telomerase reverse transcriptase (hTERT) antigen and in vitro bioimaging application

Abstract

Lung cancer (LC) is a deadly malignancy that is posing a serious threat to human health. Therefore, early detection of LC biomarkers is the key to reducing LC-related fatalities. Herein, we present the first fluorescent-based selective detection of LC biomarker human telomerase reverse transcriptase (hTERT) using polyethyleneimine (PEI) functionalized graphene quantum dots (fGQDs). One-pot in situ synthesis of amine-functionalized GQDs was accomplished by hydrothermal carbonization of biowaste-derived cellulose and PEI. Synthesized fGQDs were characterized by various analytical techniques. Synthesized fGQDs not only exhibited enhanced fluorescence life-time but also excellent stability in the different solvents compared to bare GQDs. The surface activation of hTERT-Ab by carbodiimide chemistry (EDC-NHS) resulted in stacking interactions with fGQDs, involving adsorption-desorption as well as competitive mechanisms. The higher inherent affinity of hTERT-Ag (hTERT antigen) for hTERT-Ab (hTERT antibody) resulted in complex formation and recovery of fGQD fluorescence. As a result, this fluorescence sensing demonstrated a greater linear detection range (0.01 ng mL-1 to 100 µg mL-1) as well as a notable low detection limit (36.3 pg mL-1). Furthermore, the fabricated immunosensor (Ab@fGQDs) has excellent stability and performance in real samples, with an average recovery of 97.32%. The results of cytotoxicity and cellular bioimaging study in A549 cells show that fGQDs can be used for additional nanotherapeutics and biological applications.