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.
Highlights
Neurological degenerative disorders are a frightening health burden
Since the the gold gold film film electrodes electrodes used used in maintain the integrity of the biosensor prototype, modifications of the pretreatment procedure were maintain the integrity of the biosensor prototype, modifications of the pretreatment procedure were needed in in comparison comparison to to the the bulk bulk gold gold nanoparticles nanoparticles or or to to the the treated treated particles
In order to validate the enhancement of the sensor response, reproducibility, and electron charge transfer due to the cleaning procedure, electrochemical impedance spectroscopy (EIS) was employed for two groups of sensors consisting of four sensors each
Summary
Neurological degenerative disorders are a frightening health burden. These disorders include. Diagnostic analysis of T-Tau protein in blood samples on brain injury in concussed ice hockey players by the Simoa technique has been reported [16] This detection was very time consuming, and the blood samples were collected at 1, 12, 36, and 48 h and collected in an ethlenediaminetetracetic acid (EDTA) tube, and an immuno-assay was employed in the blood sample analysis. The development of a cost-effective, single-use, disposable in vitro biosensor system which can measure T-Tau in blood accurately can be a first step in providing a practical and useful tool in the assessment of neuro-degenerative disorders, and it is the objective of this study. The results of this study suggested that a useful tool for the single-use, disposable in vitro measurement of T-Tau to assess neuro-degenerative disorders including Alzheimer’s disease, TBI, and other dementia symptoms in blood serum was feasible. The level of 1400 pg/mL of T-Tau in a physiological fluid is used as the guide for identifying neuro-degenerative disorders
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