title: A Single-Use, In Vitro Biosensor for the Detection of T-Tau Protein, A Biomarker of Neuro-Degenerative Disorders, in PBS and Human Serum Using Differential Pulse Voltammetry (DPV).
abstract: A single-use, in vitro biosensor for the detection of T-Tau protein in phosphate-buffer saline (PBS) and undiluted human serum was designed, manufactured, and tested. Differential pulse voltammetry (DPV) served as the transduction mechanism. This biosensor consisted of three electrodes: working, counter, and reference electrodes fabricated on a PET sheet. Both working and counter electrodes were thin gold film, 10 nm in thickness. Laser ablation technique was used to define the size and structure of the biosensor. The biosensor was produced using cost-effective roll-to-roll process. Self-assembled monolayers (SAM) of 3-mercaptopropionic acid (MPA) were employed to covalently immobilize the anti-T-Tau (T-Tau antibody) on the gold working electrode. A carbodiimide conjugation approach using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) cross-linked anti-T-Tau to the carboxylic groups on one end of the MPA. A T-Tau protein ladder with six isoforms was used in this study. The anti-T-Tau concentration used was 500,000 pg/mL. The T-Tau protein concentration ranged from 1000 pg/mL to 100,000 pg/mL. DPV measurements showed excellent responses, with a good calibration curve. Thus, a practical tool for simple detection of T-Tau protein, a biomarker of neuro-degenerative disorders, has been successfully developed. This tool could also be extended to detect other biomarkers for neuro-degenerative disorders, such as P-Tau protein and β-amyloid 42.
authors.affiliation: Department of Chemical & Biomolecular Engineering and Electronics Design Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA. yxd176@case.edu.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Dai
authors.affiliation: Dip. Chimica Materiali e Ing. Chimica "Giulio Natta", Politecnico di Milano, Via Mancinelli 7, 20131, Italy. axm1058@case.edu.
authors.forename: Alireza
authors.initials: A
authors.lastname: Molazemhosseini
authors.affiliation: Department of Chemical & Biomolecular Engineering and Electronics Design Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA. cxl9@case.edu.
authors.forename: Chung Chiun
authors.initials: CC
authors.lastname: Liu
mesh_terms: D015415:Biomarkers / Q000032:analysis* / Q000097:blood; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002021:Buffers; D055664:Electrochemical Techniques / Q000295:instrumentation; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D057230:Limit of Detection; D019636:Neurodegenerative Diseases / Q000097:blood / Q000175:diagnosis*; D010316:Particle Size; D016875:tau Proteins / Q000032:analysis* / Q000097:blood
chemical_list: D015415:Biomarkers; D002021:Buffers; C054369:MAPT protein, human; D016875:tau Proteins; D006046:Gold
keywords: 3-mercaptopropionic acid (MPA); T-Tau protein detection; [Fe(CN)6]3−/4− redox probe; differential pulse voltammetry