Rapid electrokinetic detection of low-molecular-weight thiols by redox regulatory protein-DNA interaction in microfluidics

Lee, Jin Oh; Choi, Nakchul; Song, Simon; Kim, Young‐Pil

doi:10.1016/j.snb.2021.129735

Cited by 3 publications

(2 citation statements)

References 40 publications

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“…During the past years, the principle of electrokinetic-biosensing, exploiting streaming current/potential, has been applied in the detection of a wide variety of bio-analytes including proteins and ligands [1][2][3], DNA [4], and extracellular vesicles (EVs) [5], thereby demonstrating a potential of the method as a generic biosensor approach. Electrokinetic biosensing relies on the electrostatic and hydrodynamic interaction at the solid-liquid interface inside a microchannel and allows for label-free detection of bio-analytes.…”

Section: Introductionmentioning

confidence: 99%

Design and optimization of silicon-based electrokinetic microchip for sensitive detection of small extracellular vesicles

Gevari

Sahu

Mitra

et al. 2022

Preprint

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In this study, we demonstrate the fabrication, characterization, and application of a microchip-based electrokinetic biosensor which exploits streaming current for signal transduction. The sensor chips, fabricated using standard silicon fabrication techniques, are based on Si-glass microfluidics offering precision, robustness, and transparency. A custom-built chip-manifold allowing easy interfacing with standard microfluidic connections is also developed which shows leak-free integration of the microchips up to 6 bar of applied pressure. Within this range of pressure, the devices show linear and highly reproducible values for flow rates and streaming current with RMS noise below 20 pA. The microchips designed for multiplexed measurements were tested with the detection of free proteins (streptavidin) and also transmembrane proteins of small extracellular vesicles (sEVs) to demonstrate the capacity of the microchips to detect various types of bio-analytes. The limit of detection (LOD) for streptavidin was estimated to be 0.5 nM while for the transmembrane protein (CD9), the LOD was found to be 1.2e6 sEVs/mL. The sensitivity (LOD) of the devices was found to be about 4 times better in targeting CD9 transmembrane protein of H1975 extracellular vesicles when compared to commercial silica capillary which was used previously. The improvement in LOD is attributed to the higher surface quality of the sensor in terms of the density of surface charges which may be further exploited for even lower LOD. In addition, optical detection of fluorophore-tagged standard proteins was done through the optical window of the chip manifold and the transparent glass cap of the microchip.

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Section: Introductionmentioning

confidence: 99%

Design and optimization of silicon-based electrokinetic microchip for sensitive detection of small extracellular vesicles

Gevari

Sahu

Mitra

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…During the past years, the principle of electrokinetic biosensing, exploiting streaming current/potential, has been applied in the detection of a wide variety of bioanalytes including proteins, − DNA, and extracellular vesicles (EVs), thereby demonstrating a potential of the method as a generic biosensor approach. Electrokinetic biosensing relies on the electrostatic and hydrodynamic interaction at the solid–liquid interface inside a microchannel and allows for label-free detection of bioanalytes.…”

mentioning

confidence: 99%

Design and Optimization of a Silicon-Based Electrokinetic Microchip for Sensitive Detection of Small Extracellular Vesicles

Talebian Gevari,

Sahu,

Stridfeldt

et al. 2024

ACS Sens.

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Detection of analytes using streaming current has previously been explored using both experimental approaches and theoretical analyses of such data. However, further developments are needed for establishing a viable microchip that can be exploited to deliver a sensitive, robust, and scalable biosensor device. In this study, we demonstrated the fabrication of such a device on silicon wafer using a scalable silicon microfabrication technology followed by characterization and optimization of this sensor for detection of small extracellular vesicles (sEVs) with sizes in the range of 30 to 200 nm, as determined by nanoparticle tracking analyses. We showed that the sensitivity of the devices, assessed by a common protein−ligand pair and sEVs, significantly outperforms previous approaches using the same principle. Two versions of the microchips, denoted as enclosed and removable-top microchips, were developed and compared, aiming to discern the importance of high-pressure measurement versus easier and better surface preparation capacity. A custom-built chip manifold allowing easy interfacing with standard microfluidic connections was also constructed. By investigating different electrical, fluidic, morphological, and fluorescence measurements, we show that while the enclosed microchip with its robust glass-silicon bonding can withstand higher pressure and thus generate higher streaming current, the removable-top configuration offers several practical benefits, including easy surface preparation, uniform probe conjugation, and improvement in the limit of detection (LoD). We further compared two common surface functionalization strategies and showed that the developed microchip can achieve both high sensitivity for membrane protein profiling and low LoD for detection of sEV detection. At the optimum working condition, we demonstrated that the microchip could detect sEVs reaching an LoD of 10 4 sEVs/ mL (when captured by membrane-sensing peptide (MSP) probes), which is among the lowest in the so far reported microchipbased methods.

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Study the Profile of Some Antioxidant Markers in Diabetic Mellitus and Non-Diabetic Patients with Cardiovascular Disease

Zainal

2022

Medical Journal of Babylon

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Background: Cardiovascular diseases (CVDs) claim the lives of millions worldwide. Objective: Evaluate the levels of some antioxidant biomarkers in sera of patients with CVDs with and without diabetes mellitus type 2. Materials and Methods: Includes 100 samples in 68 patients (40 with CVDs and 28 CVDs with diabetic), and 32 healthy subjects. Results: Serum (Albumin, thiol, thiol/T.protein, amine, free amine/ T.protein, carbonyl, carbonyl/ T.protein, and Ischemia modified albumin (IMA)) showed a stronger response, a significant rise in carbonyl and carbonyl/TP, and a significant reduction in thiol and thiol/TP in the patients with CVDs with T2D compared to CVDs may be a good factor to differentiate them or predict more serious complications. Conclusions: Oxidation markers may be useful in monitoring CVDs with and without diabetes.

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Rapid electrokinetic detection of low-molecular-weight thiols by redox regulatory protein-DNA interaction in microfluidics

Cited by 3 publications

References 40 publications

Design and optimization of silicon-based electrokinetic microchip for sensitive detection of small extracellular vesicles

Design and optimization of silicon-based electrokinetic microchip for sensitive detection of small extracellular vesicles

Design and Optimization of a Silicon-Based Electrokinetic Microchip for Sensitive Detection of Small Extracellular Vesicles

Study the Profile of Some Antioxidant Markers in Diabetic Mellitus and Non-Diabetic Patients with Cardiovascular Disease

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