Detection Of Tau Protein Research Articles

Loach Mucus-Like Guanosine-Based Hydrogel as Antifouling Coating for Electrochemical Detection of Tau Protein

Loach Mucus-Like Guanosine-Based Hydrogel as Antifouling Coating for Electrochemical Detection of Tau Protein

Monitoring Protein Biochemistry on Surfaces By Electrochemical Methods

Tau protein is a neuronal protein which undergoes modifications, such as amino acid phosphorylation, as well as self-assembly into cytotoxic structures. This process leads to neurodegeneration which remains without a differential diagnosis and cure. Hence, tau protein is a viable biomarker of diseases, such as Alzheimer’s disease and other tauopathies. We have used electrochemical methods to explore various aspects of tau biochemistry: 1) interactions with binding partners, 2) kinase-catalyzed phosphorylations, 3) kinase-inhibition, and 4) biomarker detection. Specifically, by using electrochemical impedance spectroscopy, we have developed immunoassay for detection of tau protein. In addition, we screened drug targets, such as antibodies, as potential immunotherapies targeting tau. The fundamental binding studies between tau protein and other biologically-relevant binding partners, such as ferritin and transferrin, were also conducted and will be described. Our work illustrates that electrochemical methodologies are valuable tools for evaluation of protein biochemistry, and may lead to early detection, differential diagnosis and identification of viable drug therapies.

Novel Electrochemical Molecularly Imprinted Polymer-Based Biosensor for Tau Protein Detection

A novel electrochemical biosensor based on a molecularly imprinted polymer (MIP) was developed for the impedimetric determination of Tau protein, a biomarker of Alzheimer’s disease (AD). Indeed, a recent correlation between AD symptoms and the presence of Tau proteins in their aggregated form made hyperphosphorylated Tau protein (Tangles) a promising biomarker for Alzheimer’s diagnosis. The MIP was directly assembled on a screen-printed carbon electrode (C-SPE) and prepared by electropolymerization of 3-aminophenol (AMP) in the presence of the protein template (p-Tau-441) using cyclic voltammetry. The p-Tau-441 protein bound to the polymeric backbone was digested by the action of the proteolytic activity of proteinase K in urea and then washed away to create vacant sites. The performances of the corresponding imprinted and non-imprinted electrodes were evaluated by electrochemical impedance spectroscopy. The detection limit of the MIP-based sensors was 0.02 pM in PBS buffer pH 5.6. Good selectivity and good results in serum samples were obtained with the developed platform. The biosensor described in this work is a potential tool for screening Tau protein on-site and an attractive complement to clinically established methodologies methods as it is easy to fabricate, has a short response time and is inexpensive.

Bioelectrochemistry of Neuronal Protein Tauoral

Tau is a neuronal protein and one of the biomarkers of neurodegenerative diseases, such as Alzheimer’s disease. Tau protein undergoes post-translational modifications, aggregation, and is a viable drug target. In addition, this protein is a vital biomarker in biological fluids towards early detection of neurodegenerative diseases. We reported on using electrochemical impedance spectroscopy and cyclic voltammetry for detection of protein-protein interactions (1), protein-ligand interactions (2), enzymatic catalysis and enzyme inhibition (3) while focusing on tau protein and its biochemistry. For example, tau-tau, tau-ferritin, tau-transferring protein interactions were monitored, and protein kinase-catalyzed phosphorylation of tau protein was detected. The phosphorylation inhibitors, such as antibodies, were screened for their efficacies. New research directions in neuroelectrochemistry of tau protein alongside other neuronal biomarkers, such as Amyloid beta and alpha synuclein, will also be described. References A) Carlin, N., Martic-Milne, S. (2018). Anti-tau antibodies based electrochemical sensor for detection of tau protein. J. Electrochem. Soc.165: G3018-G3025. B) Jahshan, A., Esteves, J.O.V., Martic-Milne, S. (2016). Evaluation of ferritin and transferrin binding to tau protein. J. Inorg. Biochem. 162: 127-134. C) Esteves, J.O.V., Trzeciakiewicz, H., Loeffler, D.A., Martic, S. (2015). Effects of tau domain-specific antibodies and intravenous immunoglobulin on tau aggregation and aggregate degradation. Biochemistry. 54: 15-18.Trzeciakiewicz, H., Esteves, J.O.V., Carlin, N., Martic, S. (2015). Electrochemistry of heparin binding to tau protein on Au surfaces. Electrochim. Acta. 162: 24-30,Esteves, J.O.V., Martic-Milne, S. (2016). Electrochemical detection of anti-tau antibodies binding to tau protein and inhibition of GSK-3-β-catalyzed phosphorylation. Anal. Biochem. 496: 55-62

Polyvalent Biotinylated Aptamer Scaffold for Rapid and Sensitive Detection of Tau Proteins.

Biomimetic construction of artificial scaffolds has attracted increasing attention. However, the construction methods usually require redundant materials and procedures, which is inconvenient for further application. Herein, inspired by the polyvalent multifunctional structure in nature, we have designed a polyvalent biotinylated aptamer scaffold (PBAS) which can conduct analytical performance with high sensitivity and simplified procedures. To construct a PBAS, the aptamers are designed to hybridize with prepared linker probes to form polyvalent biotinylated scaffolds, which contain both multiple aptamers and signal labels. Therefore, multifunctional scaffolds can be constructed with high recognition and capture efficiency as well as significant signal amplification. Furthermore, the scaffold can be used for the assay of some disease marker proteins. By taking tau proteins as an example, the proposed aptasensor can exhibit excellent performance with a low detection limit of 153 pg mL-1 and a short assay time of 50 min, which is much better than most of the previous methods. By assays of tau proteins in both serum and artificial cerebro spinal fluid, the PBAS-based aptasensor can work well. Therefore, the scaffold may be expected to be a powerful analytical tool which may have wide applications in the detection of a variety of analytes.

A transistor-based label-free immunosensor for rapid detection of tau protein

Tau protein in cerebrospinal fluid (CSF) is a central and relevant biomarker of Alzheimer’s disease (AD) that correlates with the severity of dementia. Unfortunately, so far, direct label-free detection of tau remains a challenge. Here, we present a transistor-based biosensor that detects the net charge of tau protein directly under physiological conditions. To achieve this, readily available whole anti-tau IgG antibodies are co-immobilized on the sensor surface with polyethylene glycol (PEG) molecules of different molecular weight. We show that by increasing the PEG size from 10 kDa to 20 kDa, the electrical response upon binding of tau improves significantly. These results support recent theoretical work that predicted larger PEGs to form a thicker surface layer with a higher detectable analyte charge. With 20 kDa PEG, we demonstrate label-free tau detection in a wide concentration range with detection limits <1 pM in 150 mM buffer and cell culture media, as well as < 10 pM in artificial CSF. This purely electrical method allows fast and simple tau detection within 30 min without sample processing, washing steps, or labeled detection antibodies. By exchanging the capture antibody, the platform is also amenable to different biomarkers and may enable future diagnostic tools for AD and other diseases.

A dip-and-read optical aptasensor for detection of tau protein.

Neurodegeneration currently remains without a differential diagnosis or cure. Tau protein is one of the biomarkers of neurodegenerative diseases commonly known as tauopathies. Tau protein plays an integral role in stabilizing microtubules and cell structure; however, due to post-translational modifications, tau protein undergoes self-assembly into cytotoxic structures and is co-localized intra- and extracellularly. Hence, tau protein is a viable biomarker associated with protein pathogenesis and neurodegeneration. The novel optical biosensor for tau441 protein is based on the aptamer recognition probe and the biolayer interferometry (BLI) method for detection. The current biotin-aptasensor in combination with the streptavidin surface provides real-time monitoring of tau441 protein in the nanomolar range, with the limit of detection at 6.7nM in vitro. The tau441 detection is achieved with high selectivity over other neurodegeneration biomarkers which include amyloid-β and α-synuclein. The aptasensor also allows for tau441 protein detection in a complex matrix such as fetal bovine serum, indicating its utility in other biological fluids for diagnostic applications. The optical method is simple, rapid and highly selective for point-of-care application which is critical for achieving the early and differential diagnosis of neurodegenerative diseases and identifying their treatments. Graphical abstract.

Immunoglobulin G-Based Steric Hindrance Assay for Protein Detection.

With the imminent needs of rapid, accurate, simple point-of-care systems for global healthcare industry, electrochemical biosensors have been widely developed owing to their cost-effectiveness and simple instrumentation. However, typical electrochemical biosensors for direct analysis of proteins in the human biological sample still suffer from complex biosensor fabrication, lack of general method, limited sensitivity, and matrix-caused biofouling effect. To resolve these challenges, we developed a general electrochemical sensing strategy based on a designed steric hindrance effect on an antibody surface layer. This strategy utilizes the interaction pattern of protein-G and immunoglobulin G (Fc and Fab regions), providing a steric hindrance effect during the target capturing process. The provided steric hindrance effect minimizes the matrix effect-caused fouling surface and altered the path of electron transfer, delivering a low-fouling and high-sensitivity detection of protein in complex matrices. Also, an enzyme-based horseradish peroxidase/hydroquinone/H2O2 transduction system can also be applied to the system, demonstrating the versatility of this sensing strategy for general electrochemical sensing applications. We demonstrated this platform through the detection of Tau protein and programming death ligand 1 with a subpico molar detection limit within 10 min, satisfying the clinical point-of-care requirements for rapid turnaround time and ultrasensitivity.

Step-by-step monitoring of a magnetic and SERS-active immunosensor assembly for purification and detection of tau protein.

We report a bottom‐up synthesis of iron oxide and gold nanoparticles, which are functionalized and combined to form a nanohybrid serving as an immune sensor, which selectively binds to tau protein, a biomarker for diagnosis of Alzheimer's disease. Detection of the target analyte is achieved by surface‐enhanced Raman scattering originating from the diagnostic part of the nanohybrid that was prepared from Au nanoparticles functionalized with 5,5′‐dithiobis‐(2‐nitrobenzoic acid) as a Raman reporter and monoclonal anti‐tau antibody. The magnetic part consists of FexOy nanoparticles functionalized with polyclonal anti‐tau antibody and is capable to separate tau protein from a complex matrix such as cerebrospinal fluid. We further identified and validated a set of analytical tools that allow monitoring the success of both nanoparticle preparation and each functionalization step performed during the assembly of the two binding sites by an immune reaction. By applying UV/Vis spectroscopy, dynamic light scattering, zeta potential measurements, X‐ray diffraction, small‐angle X‐ray scattering, and transmission electron microscopy, we demonstrate a proof‐of‐concept for a controlled and step‐by‐step traceable synthesis of a tau protein‐specific immune sensor.

Robust and Universal SERS Sensing Platform for Multiplexed Detection of Alzheimer's Disease Core Biomarkers Using PAapt-AuNPs Conjugates.

Multiplexed detection of Alzheimer's disease (AD) core biomarkers is of great significance to early diagnosis and personalized treatment of AD patients. Herein, we construct a robust and convenient surface-enhanced Raman scattering (SERS) biosensing platform for simultaneous detection of Aβ(1-42) oligomers and Tau protein using different Raman dye-coded polyA aptamer-AuNPs (PAapt-AuNPs) conjugates. This strategy relies on the specific protein-aptamer binding-mediated aggregation of AuNPs and the concomitant plasmonic coupling effect that allow us to "turn on" SERS detection of protein biomarkers. To the best of our knowledge, this is the first work in which PAapt-AuNPs conjugates are used for probing protein biomarkers, which may be enlightening for the exploitation of more extensive biological applications of aptamer-AuNPs conjugates. The results reveal that the present strategy displays excellent analytical performance. Moreover, the applicability of this strategy is demonstrated in the artificial cerebrospinal fluid (CSF) samples with satisfactory results. Except for the prominent sensitivity and practicality, our strategy offers additional advantages. The preparation of nanoconjugates is handy and easily repeated, and the synthesis cost is greatly reduced because it dispenses with the complicated labeling process. Moreover, the assay can be accomplished in 15 min, allowing rapid detection of protein biomarkers. Furthermore, simultaneous detection of Tau protein and Aβ(1-42) oligomers is realized by employing different Raman dye-coded nanoconjugates, which is valuable for accurately predicting and diagnosing AD disease. Thus, our PAapt-AuNPs conjugate-based multiplexed SERS strategy indeed creates a useful and universal platform for detecting multiple protein biomarkers and related clinical diagnosis.

A tyrosinase-induced fluorescence immunoassay for detection of tau protein using dopamine-functionalized CuInS2/ZnS quantum dots.

Rapid, highly sensitive detection of tau protein and other neurodegenerative biomarkers remains a significant hurdle for diagnostic tests for Alzheimer's disease. In this work, we developed a novel tyrosinase (TYR)-induced tau aptamer-tau-tau antibody (anti-tau) sandwich fluorescence immunoassay to detect tau protein that used dopamine (DA)-functionalized CuInS2/ZnS quantum dots as the fluorophore. CuInS2/ZnS core/shell quantum dots with high luminescence, low toxicity, and excellent biocompatibility were successfully fabricated and decorated with DA through amide conjugation. Meanwhile, TYR was conjugated with anti-tau by a click reaction. When DA-functionalized CuInS2/ZnS quantum dots were added to the sandwich system, TYR catalyzed the transformation of DA to dopamine quinone, which acted as an effective electron acceptor and triggered fluorescence quenching. The fluorescence intensity of the immunoassay based on DA-functionalized CuInS2/ZnS quantum dots shows good performance in terms of linearity with the logarithm of tau protein concentration, with a linear concentration range from 10 pM to 200 nM. This work is the first to use a TYR-induced fluorescence immunoassay for the rapid detection of tau protein, paving a new way for the detection of disease biomarkers. Graphical abstract.

Improving Sensitivity and Specificity of Amyloid-β Peptides and Tau Protein Detection with Antibiofouling Magnetic Nanoparticles for Liquid Biopsy of Alzheimer's Disease.

Alzheimer's disease (AD) is a growing global healthcare burden affecting the aging population and society. Given the lack of effective treatment to AD, early detection at the prodromal stage and timely monitoring of changes during progression are considered the best approach to control and intervene in disease progression. "Liquid biopsy" of AD biomarkers amyloid-β peptides (Aβs) and tau proteins in the cerebrospinal fluid (CSF) or blood samples holds great promises for cost-effective, widely accessible, and easy-administrated noninvasive detection and follow-up of AD. However, current in vitro detection methods have not yet demonstrated sufficient sensitivity and specificity using neither Aβs nor tau proteins biomarkers. One major challenge of accurate detection and measurement of biomarker levels in biofluidic samples is the biofouling effect with nonspecific adsorption of unwanted biomolecules, such as various serum proteins, on the surface of targeted detecting agents or devices, causing false-positive and false-negative findings. In this study, antibiofouling polymer polyethylene glycol-block-allyl glycidyl ether (PEG-b-AGE) coated magnetic iron oxide nanoparticles (IONPs) capable of suppressing the nonspecific interactions with biomolecules, especially proteins, were investigated for the immunomagnetic capturing of Aβ40 and Aβ42 peptides and tau protein spiked in CSF- and serum-mimicking samples using corresponding antibodies conjugated as targeting ligands. Antibody-conjugated antibiofouling IONPs demonstrated improved specificity (>90%) and sensitivity (>95%) over those of antibody-conjugated magnetic micron beads (Dynabeads, ∼50% specificity and 30-40% sensitivity) widely used as magnetic separating agents under the same experimental conditions with the presence of nontargeted interfering proteins. The antibody-conjugated IONPs also exhibited significantly higher sensitivities (80-90%) and better performance of capturing Aβs and tau protein from the human whole blood samples than antibody-conjugated Dynabeads (∼20%).

Controlled fluorescence quenching by antibody-conjugated graphene oxide to measure tau protein.

We report an ultrasensitive immunoassay for tau protein—a key marker of Alzheimer's disease. This sensing platform relies on graphene oxide (GO) surfaces conjugated with anti-human tau antibody to provide quantitative binding sites for the tau protein. The GO quenches standard fluorescein isothiocyanate labelled tau (tau-FITC) when tau protein and tau-FITC are both present and compete for the binding sites. This change in fluorescence signal can be used to quantitate tau protein. In contrast with traditional enzyme-linked immunosorbent assay (ELISA), our method does not require enzyme-linked secondary antibodies for protein recognition nor does it require an enzyme substrate for optical signal generation. This requires fewer reagents and has less systematic error than the antigen–antibody recognition steps in ELISA. Our method has a tau protein detection limit of 0.14 pmol ml−1 in buffer. This approach could be developed into a promising biosensor for the detection of tau protein and may be useful in the clinical diagnosis of tau-induced neurodegeneration syndromes.

Real-Time Tau Protein Detection by Sandwich-Based Piezoelectric Biosensing: Exploring Tubulin as a Mass Enhancer.

Human tau protein is one of the most advanced and accepted biomarkers for AD and tauopathies diagnosis in general. In this work, a quartz crystal balance (QCM) immunosensor was developed for the detection of human tau protein in buffer and artificial cerebrospinal fluid (aCSF), through both direct and sandwich assays. Starting from a conventional immuno-based sandwich strategy, two monoclonal antibodies recognizing different epitopes of tau protein were used, achieving a detection limit for the direct assay in nanomolar range both in HBES-EP and aCSF. Afterward, for exploring alternative specific receptors as secondary recognition elements for tau protein biosensing, we tested tubulin and compared its behavior to a conventional secondary antibody in the sandwich assay. Tau–tubulin binding has shown an extended working range coupled to a signal improvement in comparison with the conventional secondary antibody-based approach, showing a dose–response trend at lower tau concentration than is usually investigated and closer to the physiological levels in the reference matrix for protein tau biomarker. Our results open up new and encouraging perspectives for the use of tubulin as an alternative receptor for tau protein with interesting features due to the possibility of taking advantage of its polymerization and reversible binding to this key hallmark of Alzheimer’s disease.

ЕКСПРЕСІЯ TAU-БІЛКУ В СЕНСОМОТОРНІЙ КОРІ ПРИ МОДЕЛЮВАННІ ТРАНЗИТОРНОЇ ІШЕМІЇ ТА ІМУНОКОРЕКЦІЇ

Вступ. Ішемія головного мозку супроводжується дифузним пошкодженням структурних елементів нейронів. Одним із маркерів, який демонструє функціональні процеси в нервовій системі, є tau -білок.Мета дослідження – вивчити особливості експресії tau-білку в сенсомоторній корі за умов порушення кровопостачання та корекції нейродегенеративних змін імуномодуляцією.Матеріали і методи. При моделюванні транзиторної ішемії та корекції нейродегенеративних змін імунофаном, був проведений експеримент на 150 білих статевозрілих щурах-самцях масою 260–290 г. Були застосовані гістологічні, імуногістохімічний, морфометричний та статистичний методи дослідження.Результати. Проведені спостереження показали, що порушення кровообігу у корі мозку призводять до зростання експресії tau-білку в сенсомоторній корі. Виразність цих змін залежить від ступеню порушення кровообігу, що непрямим шляхом підтверджується наявністю змін експресії tau-білку в контрлатеральній півкулі.Стрімке наростання експресії tau-білку в гостру фазу після ішемічної атаки свідчить про дезорганізацію цитоскелету нервових волокон та, певною мірою, відображує явища аутонейротомії. Поява tau-білку в перикаріонах нейроцитів може бути показником тяжкості дегенеративних процесів. Виявлення tau-білку в тілах гліоцитів може бути розцінені як явища фагоцитозу продуктів розпаду.Застосування імунофану, який має певний нейропротекторний ефект, показало його здатність зменшувати індуковане ішемією підвищення експресії tau-білку. Це може бути пояснено як імунорегулюючими властивостями цього пептиду (аргініл-альфа-аспартил-лізил-валіл-тирозил-аргінін), так і його детоксикаційною дією, блокуванням вільнорадикальних процесів перекисного окиснення, запобіганням пошкодженню лімфоцитів та гранулоцитів, що приймають участь у нейрозапеленні .Висновки. Транзиторне порушення кровопостачання сенсомоторної кори великих півкуль призводить до зростання в ній експресії tau-білку, повного відновлення якої не відбувається навіть через 3 місяці після ішемічної атаки. Дисциркуляторні зміни у мозку, що виникають при мобілізації або перев’язуванні сонної артерії з боку операції, а також ті що виникають у контрлатеральній півкулі мозку, призводять до незначного, але на певному етапі, статистично значущому підвищені експресії tau-білку.Застосування імунофану при порушенні кровопостачання мозку призводить до менш виразного зростання експресії tau-білку в сенсомоторній корі.

Anti-Tau Antibodies Based Electrochemical Sensor for Detection of Tau Protein Biomarkers

Neuronal tau protein is one of the biomarkers of neurodegeneration which remains without a cure and differential diagnosis. Hence, novel methods are needed for detection of tau protein. Toward this aim, the detection of full-length tau441 protein was achieved using the electrochemical immunoassay based on direct immobilization of anti-tau antibodies on Au surface by using an active N-hydroxysuccinimide ester coupling chemistry (Ab-Au). The electrochemical impedance spectroscopy was used to monitor tau441 binding to Ab-Au in the presence [Fe(CN)6]3−/4− redox probe. The reduction in the charge-transfer resistance, Rct, was observed upon tau441 binding to Ab-Au. Detection of tau441 was observed in μM to nM range with high specificity and selectivity. The electrochemical immunosensor exhibited minimal non-specific binding to bovine serum albumin. Tau441 did not bind to Au surface functionalized with non-tau-specific antibody MOPC-21. The electrochemical biosensor was optimized by screening various anti-tau antibodies. The tau441 binding to anti-tau antibodies was also measured using Enzyme-Linked Immunosorbent Assay (ELISA). The label-free electrochemical impedance spectroscopy immunosensor may be used for selective detection of tau protein in biological fluids, as a biomarker of neurodegeneration.

Detection of the tau protein in human serum by a sensitive four-electrode electrochemical biosensor

This study presents a novel approach based on a four-electrode electrochemical biosensor for the detection of tau protein – one of the possible markers for the prediction of Alzheimer's disease (AD). The biosensor is based on the formation of stable antibody–antigen complexes on gold microband electrodes covered with a layer of a self-assembled monolayer and protein G. Antibodies were immobilized on the gold electrode surface in an optimal orientation by protein G interaction. Electrochemical impedance spectroscopy was used to analyze impedance change, which revealed a linear response with increasing tau concentrations. The assay is fast (<1h for incubation and measurement) and very sensitive. The limit of quantification for the full-length 2N4R tau protein is 0.03pM, a value unaltered when the assay was processed in bovine serum albumin or human serum. This technology could be adapted for the detection of other biomarkers to provide a multiple assay to identify AD progression in a point of care setting.

Toward sensitive immuno-based detection of tau protein by surface plasmon resonance coupled to carbon nanostructures as signal amplifiers

Interest on Tau protein is fast increasing in Alzheimer's disease (AD) diagnosis. There is the urgent need of highly sensitive and specific diagnostic platforms for its quantification, also in combination with the other AD hallmarks. Up to now, SPR has been poorly exploited for tau detection by immunosensing, due to sensitivity limits at nanomolar level, whereas the clinical requirement is in the picomolar range. Molecular architectures built in a layer-by-layer fashion, biomolecules and nanostructures (metallic or not) may amplify the SPR signal and improve the limit of detection to the desired sensitivity. Mostly gold nanostructures are widely employed to this aim, but great interest is also emerging in Multi Walled Carbon Nanotubes (MWCNTs). Here MWCNTs are modified and then decorated with the secondary antibody for tau protein. Eventually we took advantage from MWCNTs-antibody conjugate to obtain a sandwich-based bioassay with the capability to increase the SPR signal of about 102 folds compared to direct detection and conventional unconjugated sandwich. With respect to these results, we hope to give a strong impulse for further investigation on studying possible roles of carbon nanotubes in optical-based biosensing.

Femtomolar Detection of Tau Proteins in Undiluted Plasma Using Surface Plasmon Resonance.

The ability to directly detect Tau protein and other neurodegenerative biomarkers in human plasma at clinically relevant concentrations continues to be a significant hurdle for the establishment of diagnostic tests for Alzheimer's disease (AD). In this article, we introduce a new DNA aptamer/antibody sandwich assay pairing and apply it for the detection of human Tau 381 in undiluted plasma at concentrations as low as 10 fM. This was achieved on a multichannel surface plasmon resonance (SPR) platform with the challenge of working in plasma overcome through the development of a tailored mixed monolayer surface chemistry. In addition, a robust methodology was developed involving various same chip control measurements on reference channels to which the detection signal was normalized. Comparative measurements in plasma between SPR and enzyme-linked immunosorbent assay (ELISA) measurements were also performed to highlight both the 1000-fold performance enhancement of SPR and the ability to measure both spiked and native concentrations that are not achievable with ELISA.

Microtubule density and landing rate as parameters to analyze tau protein in the MT-kinesin “gliding” assay

Microtubule-associated protein (MAP) tau is a well-established hallmark of a large group of age related neurodegenerative diseases collectively called tauopathies. Under pathological conditions the equilibrium of tau binding to the MTs is perturbed, either by misregulation in the expression levels of specific tau isoforms or by MAPT gene mutations. Preclinical detection of such misregulated tau proteins in cerebrospinal fluid (CSF) is desirable for differential diagnosis and effective prognosis of neurodegeneration. Conventional tau protein detection methods utilize tau isoform-specific antibodies. Such immuno-based protocols, including enzyme-linked immunosorbent assay (ELISA) and Western blots have appropriate sensitivity and specificity, but often show high variability and are time consuming. Here, we established a non-immuno tau protein detection method utilizing microtubule (MT)-kinesin “gliding”assay. All the six tau isoforms expressed in the human brain (0N3R, 1N3R, 2N3R, 0N4R, 1N4R and 2N4R) and five MAPT gene mutants (V248L, G272V, P301L, V337M and R406W) were studied. The landing rate, binding density and gliding velocity of MTs with respect to each tau type were determined and are proposed as tau detection parameters. The detection parameters depicted the type of tau bound to the MTs. Furthermore, MT landing rate and density were found to be superior to gliding velocity in differentiating tau isoforms and mutants. The 3R vs. 4R isoforms, their admixtures, wild vs. mutant 2N4R and specific mutants were differentiated. Our data show that MT-kinesin gliding assay provides a convenient, lab-on-a-chip (LOC) compatible and antibody-free protocol for tau protein analysis.