Microfluidic biochips for simple impedimetric detection of thrombin based on label-free DNA aptamers

Lim, Taemin; Lee, Seung Yong; Yang, Jeong-Suong; Hwang, Seung Yong; Ahn, Yonghan

doi:10.1007/s13206-016-1203-7

Cited by 19 publications

(10 citation statements)

References 29 publications

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“…In previous studies, urease was immobilized on the surface of an electrode that was used for electrochemical measurements [11,17]. However, direct immobilization limits the area of urease immobilization to the area of the electrode and increases the cost of the assay, because urease must be re-immobilized each time the electrode is replaced [18]. As a better alternative, nanostructures onto which urease could be immobilized were constructed on the surface of an electrode [11,19].…”

Section: Introductionmentioning

confidence: 99%

“…The porous membranes maximized the contact area with the fluid in the flow condition, thus improving the sensitivity of the electrochemical measurement. Finally, the urease-immobilized porous membranes were inserted into a polydimethylsiloxane (PDMS) chamber to form a fluidic system [18,28,29]. The concentration of urea was then monitored using the fabricated urea biosensor in the flow condition.…”

Section: Introductionmentioning

confidence: 99%

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Parylene-Coated Polytetrafluoroethylene-Membrane-Based Portable Urea Sensor for Real-Time Monitoring of Urea in Peritoneal Dialysate

Park

Kim

et al. 2019

Sensors

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A portable urea sensor for use in fast flow conditions was fabricated using porous polytetrafluoroethylene (PTFE) membranes coated with amine-functionalized parylene, parylene-A, by vapor deposition. The urea-hydrolyzing enzyme urease was immobilized on the parylene-A-coated PTFE membranes using glutaraldehyde. The urease-immobilized membranes were assembled in a polydimethylsiloxane (PDMS) fluidic chamber, and a screen-printed carbon three-electrode system was used for electrochemical measurements. The success of urease immobilization was confirmed using scanning electron microscopy, and fourier-transform infrared spectroscopy. The optimum concentration of urease for immobilization on the parylene-A-coated PTFE membranes was determined to be 48 mg/mL, and the optimum number of membranes in the PDMS chamber was found to be eight. Using these optimized conditions, we fabricated the urea biosensor and monitored urea samples under various flow rates ranging from 0.5 to 10 mL/min in the flow condition using chronoamperometry. To test the applicability of the sensor for physiological samples, we used it for monitoring urea concentration in the waste peritoneal dialysate of a patient with chronic renal failure, at a flow rate of 0.5 mL/min. This developed urea biosensor is considered applicable for (portable) applications, such as artificial kidney systems and portable dialysis systems.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Parylene-Coated Polytetrafluoroethylene-Membrane-Based Portable Urea Sensor for Real-Time Monitoring of Urea in Peritoneal Dialysate

Park

Kim

et al. 2019

Sensors

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“…Although some authors report lower detection limits than those obtained in this work, most of these studies employ detection techniques that require highly sophisticated equipment or expensive reagents, such as labeled oligonucleotides. [50][51][52][53][54][55][56] Additionally, none of the other reported techniques in Table 2 employ whole blood as a starting sample, or uses a chimeric label-free aptamer. For the application assay we only required an oligonucleotide, simple folding conditions plus a dye that is noncovalent bound to the DNA, which guarantees the low cost of the system in comparison with labeled probes.…”

Section: Chimeric Aptafluorescence Assay Strategy Advantagesmentioning

confidence: 99%

Protein detection in blood via a chimeric aptafluorescence assay: toward point-of-care diagnostic devices

et al. 2018

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Paper-based analytics allows building portable and disposable devices for point-of-care (POC) diagnosis. Conventional methods for quantifying proteins exhibit substantial disadvantages related to costs and difficulty of the technique when used in settings where fast and cost-effective assays are needed. We report the successful application of a simple, rapid, easy to use, and label-free aptasensor strategy based on the selective fluorescence of the NMM IX dye. For the probe design, the three-dimensional (3-D) structures of the DNA components were carefully analyzed using software for the 3-D visualization of crystallographic structures. The chimeric aptafluorescence molecule consists of two modules, a detection aptamer and a transduction sequence that induces the specific fluorescence of NMM IX. In the presence of thrombin, a fluorescent spot visible to the naked eye can be observed. The fluorescent response is directly proportional to protein concentration and can be easily quantified colorimetrically using a low-cost microscopy system. The recognition probe design might be adaptable to other relevant biological analytes by changing the sequence of the aptamer. This proof of principle represents a contribution to the development of useful, cheap, reliable, and simple protein quantification assays for POC testing.

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“…The porous membranes maximized the contact area with the fluid in the flow condition, thus improving the sensitivity of the electrochemical measurement. Finally, the urease-immobilized porous membranes were inserted into a polydimethylsiloxane (PDMS) chamber to form a fluidic system [30]. The concentration of urea was then monitored using the fabricated urea biosensor in the flow condition.…”

Section: Introductionmentioning

confidence: 99%

Parylene-Coated PTFE-Membrane-Based Portable Urea Sensor for Use in Fast Flow Conditions

Park¹,

Kim²,

Kim³

et al. 2019

Preprint

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A portable urea sensor for use in the fast flow condition was fabricated using porous polytetrafluoroethylene (PTFE) membranes coated with amine-functionalized parylene, parylene-A, by vapor deposition. To generate a specific electrochemical sensor signal from urea, the urea-hydrolyzing enzyme urease was immobilized on the parylene-A-coated PTFE membranes via chemical crosslinking using glutaraldehyde. The urease-immobilized membranes were assembled in a polydimethylsiloxane (PDMS) fluidic chamber, and a screen-printed carbon three-electrode system was used for electrochemical measurements. The success of urease immobilization was confirmed using fluorescence microscopy, scanning electron microscopy, and Fourier-transform infrared spectroscopy. The optimum concentration of urease for immobilization on the parylene-A-coated PTFE membranes was determined to be 48 mg/mL, and the optimum number of membranes in the PDMS chamber was found to be 8. Using these optimized conditions, we fabricated the urea biosensor and monitored urea samples under various flow rates ranging from 0.5 to 10 mL/min in the flow condition using chronoamperometry. To test the applicability of the sensor for physiological samples, we used it for monitoring urea concentration in the waste peritoneal dialysate of a patient with chronic renal failure, at a flow rate of 0.5 mL/min.

show abstract

Microfluidic biochips for simple impedimetric detection of thrombin based on label-free DNA aptamers

Cited by 19 publications

References 29 publications

Parylene-Coated Polytetrafluoroethylene-Membrane-Based Portable Urea Sensor for Real-Time Monitoring of Urea in Peritoneal Dialysate

Parylene-Coated Polytetrafluoroethylene-Membrane-Based Portable Urea Sensor for Real-Time Monitoring of Urea in Peritoneal Dialysate

Protein detection in blood via a chimeric aptafluorescence assay: toward point-of-care diagnostic devices

Parylene-Coated PTFE-Membrane-Based Portable Urea Sensor for Use in Fast Flow Conditions

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