title: Sensitive NADH detection in a tumorigenic cell line using a nano-biosensor based on the organic complex formation.
abstract: A robust amperometric sensor for β-nicotinamide adenine dinucleotide (NADH) detection was developed through the organic complex formation with ethylenediaminetetraacetic acid (EDTA) bonded on the polyethylenimine (PEI)/activated graphene oxide (AGO) layer. The EDTA immobilized sensor probe (GCE/AGO/PEI-EDTA) revealed a catalytic property towards NADH oxidation that allows for the highly sensitive electrochemical detection of NADH at a low oxidation potential. Surface characterization demonstrated that the negatively charged AGO acted as nanofillers in the positively charged PEI matrix through the charge interaction. The immobilization of EDTA on the polymer layer provided more surface area for NADH to interact with through the enhanced chemical interlocking between them. We observed the strong interaction between NADH and EDTA on the AGO/PEI layer using a quartz crystal microbalance (QCM), X-ray photoelectron spectroscopy (XPS), and the calculation of the minimized energy for complex formation. The dynamic range of NADH was determined to be between 0.05μM and 500μM with a detection limit (LD) of 20.0±1.1nM. The reliability of the developed sensor for biomedical applications was examined by detecting NADH in tumorigenic lung epithelial cells using the standard addition method.
authors.affiliation: Department of Chemistry and Institute of Biophysio Sensor Technology (IBST), Pusan National University, Busan 46241, South Korea.
authors.forename: Mahmood H
authors.initials: MH
authors.lastname: Akhtar
authors.affiliation: Department of Chemistry and Institute of Biophysio Sensor Technology (IBST), Pusan National University, Busan 46241, South Korea.
authors.forename: Tanveer A
authors.initials: TA
authors.lastname: Mir
authors.affiliation: Department of Chemistry and Institute of Biophysio Sensor Technology (IBST), Pusan National University, Busan 46241, South Korea.
authors.forename: N G
authors.initials: NG
authors.lastname: Gurudatt
authors.affiliation: Department of Chemistry and Institute of Biophysio Sensor Technology (IBST), Pusan National University, Busan 46241, South Korea.
authors.forename: Saeromi
authors.initials: S
authors.lastname: Chung
authors.affiliation: Department of Chemistry and Institute of Biophysio Sensor Technology (IBST), Pusan National University, Busan 46241, South Korea. Electronic address: ybshim@pusan.ac.kr.
authors.forename: Yoon-Bo
authors.initials: YB
authors.lastname: Shim
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D004492:Edetic Acid / Q000737:chemistry*; D055664:Electrochemical Techniques / Q000379:methods; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D008958:Models, Molecular; D009243:NAD / Q000032:analysis* / Q000097:blood*; D009369:Neoplasms / Q000737:chemistry; D010084:Oxidation-Reduction; D010087:Oxides / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D010087:Oxides; D009243:NAD; D006108:Graphite; D011094:Polyethyleneimine; D004492:Edetic Acid
keywords: EDTA; Human serum; NADH sensor; Polymer-GO; Tumorigenic cells