• Korean Chemical Engineering Research
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• v.61 no.3
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• pp.407-411
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• 2023

The urease is covalently immobilized on the surface of the magnetic particles to catalyze the conversion of urea to bicarbonate anion. The conversion was confirmed qualitatively using high-pressure liquid chromatography and UV/Vis spectrometry, and analyzed quantitatively with cyclic voltammetry. The amount of conversion with respect to time was measured and analyzed by the reaction rate equation to calculate a reaction rate constant of 0.0474 min-1. In the 1 to 3 cycles, a conversion percentage of over 90% was found, and it was possible to reuse the urease 8 times up to the percentage of 50%. It was also observed that the stability evaluated for storage for 30 days was maintained. As a result of this study, it can be seen that the urease covalently immobilized on the scaffold can be used for urea removal for the purpose of producing ultrapure water.

• Bulletin of the Korean Chemical Society
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• v.18 no.11
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• pp.1149-1152
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• 1997

An enzyme electrode for the amperometric measurement of urea was prepared by co-immobilizing L-glutamate dehydrogenase and urease onto an Immobilon-AV affinity membrane attached to a glassy carbon electrode. The reduced nicotinamide adenine dinucleotide(NADH) was used as the electroactive species. The electrochemical oxidation of NADH was monitored at +1.0 volt vs. Ag/AgCl. The enzyme-immobilized electrode was linear over the range of 2.0 × 10-5 to 2 × 10-4 M. The response time of the electrode was approximately 3 min. and the optimum pH of the enzyme immobilized membrane was pH 7.4-7.6 (Dulbcco's buffer solution). It was stable for at least two weeks or 50 assays. There was no interference from other physiological species, except from high levels of ascorbic acid.

• Journal of the Korean Chemical Society
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• v.36 no.3
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• pp.393-404
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• 1992

The response properties of continuous automated system using an enzyme reactor for determination of urea were investigated. The enzyme reactor was constructed to packed-bed form which filled with nylon-6 beads (42∼48 mesh), which immobilized urease with glutaraldehyde, in teflon tube (2 mm I.D., 20 cm length). The system was composed of the enzyme reactor, gas dialyzer, and tublar PVC-nonactin membrane ammonium ion-selective electrode as an indicator electrode in serial order. The response characteristics of this system were as follows. That is, the concentration range of linear response, slope of linear response, detection limit, and conversion percentage were $5.5{\times}10^{-6}$∼$2.4{\times}10^{-3}M$, 57.8 mV/decade, $1.5{\times}10^{-6}$, and 80.8%, respectively. The optimum buffer and life time of urease reactor were 0.01M Tris-HCl buffer solution (pH 7.0∼7.8) and 0.01M phosphate buffer solution (pH 6.9∼7.5) and about 150 days, respectively. And the urease reactor had no interferences of the other physiological materials.

• Journal of the Korean Chemical Society
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• v.36 no.6
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• pp.864-871
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• 1992

A preparation method and response characteristics of a urea sensor which consisted of pH-ISFET and urease-immobilized membrane were investigated. The pH-ISFET urea sensor was fabricated by immobilizing BSA and urease with glutaraldehyde on gate of the pH-ISFET. Effects of pH and concentration of working buffer and enzyme load on the pontentiometric response of the pH-ISFET urea sensor were examined. Response characteristics for the determination of urea in synthetic physiological saline solution (pH 7.4) were as follows. That is, the concentration range of linear response, slope of linear response (sensitivity), and response time were 5.0 ${\times}$ $10^{-4}$ ${\sim}$ 5.0 ${\times}$ $10^{-2}$M, 31.6 mV/decade and 4${\sim}$10 min, respectively.

• BMB Reports
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• v.30 no.5
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• pp.308-314
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• 1997

We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

• Journal of Sensor Science and Technology
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• v.20 no.6
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• pp.400-405
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• 2011

This study introduced the development of a strip type disposable enzyme-linked immunosensor platform for the detection of IgG. Strips of the strip sensor were fabricated by using commercial nitrocellulose filter membranes and a housing holder for the strips was manufactured by using a standard injection molding process for a plastic material. An IgG-urease conjugate was prepared and used for the competitive immune-binding with sample IgG. From the enzymatic reaction between the conjugated urease and urea added, ammonia was generated and caused a localized alkaline pH change on the immobilized antibody band which was coated onto the sensor strips. This pH increase subsequently caused a color change of the antibody band in the presence of a pH indicator, phenol red. Used in conjunction with a competitive immunoassay format, the intensity of the color produced is directly linked with the concentration of target analyte, IgG, and specific measurement of IgG in a lateral flow immunoassay format was achieved over the range 100 ppb to 2000 ppb IgG.

• Journal of the Korean Institute of Telematics and Electronics A
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• v.28A no.12
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• pp.46-52
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• 1991

A new signal process circuit using two ISFETs as the input devices of the MOS differential amplifier stage for an ISFET biosensor has been developed. One chip integration of the newly developed signal process circuit, ISFETs and a Pt quasi-reference electrode has been carried out according to modified LOCOS p-well CMOS process. The fabricated chip showed gains of 0.8 and 1.6, good liniarity in the input-output relationship and very small power dissipation, 4mW. The chip was applied to realize a urea sensor by forming an immobilized urease membrane, using lift-off technique. on the gate of an ISFET. The urea sensor chip showed stable responses in a wide range of urea concentrations.

• Journal of Microbiology and Biotechnology
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• v.13 no.3
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• pp.373-377
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• 2003

Various groups of industrial and agricultural pollutants (heavy metal ions, cyanides, and pesticides) can be detected by enzymes. Since heavy metal ions inhibit urease, cyanides inhibit peroxidase, organophosphorus and carbamate pesticides inhibit butyrylcholinesterase, these enzymes were co-immobilized into a bovine serum albumin gel on the surface of an ion-sensitive field effect transistor to create a bioprobe that is sensitive to the compounds mentioned above. The sensitivity of the present sensor towards KCN corresponded to $1\;\mu\textrm{M}$ with 1 min of incubation time. The detection limits for Hg(II) ions and the pesticide carbofuran were 0.1 and $0.5\;\mu\textrm{M}$, respectively, when a 10 min sensor incubation time in contaminated samples was chosen. The total time for determining the concentrations of all species mentioned did not exceed 20 min.

• Journal of the Korean Chemical Society
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• v.39 no.12
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• pp.925-931
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• 1995

Urea sensors, prepared by immobilizing urease on ammonium-selective membrane electrodes doped with nonactin, can show interference from several ionic species present in blood samples (e.g., sodium, potassium, and endogenous ammonium ions). This interference problem does not arise from the immobilized biocatalytic reaction but rather from the innate response of the base transducer to ionic species in the sample. In this work, the use of calibrators containing adequate amounts of ionic species is examined to reduce errors caused by endogenous ionic interferences with blood urea measurements. Simultaneous measurements of the interfering species with additional sensors and subsequent substractions of these values from the urea electrode signals are also described. It is shown that the use of a potassium-selective electrode with an adequate calibrator system greatly enhances the accuracy of the urea sensor measurements.

• Journal of the Korean Electrochemical Society
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• v.1 no.1
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• pp.40-44
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• 1998

The LAPS device of fast response and high sensitivity, based on electrochemical potential difference, and its system were fabricated for the precise measurement of pH changes and its characteristic were investigated. The electrostatic variation characteristics of LAPS according to the pH changes and parameters in the device were verified through a simulation using LAPS equivalent circuit model. The LAPS device and its system were fabricated on the basis of the result of simulation. The fabricated LAPS system showed linear sensitivity (about 56 mV/pH within the range of pH 2 to pH 11. In order to overcome the defect of general urea sensor (especially slow response time), urease immobilized nitrocellulose membrane was attached on the LAPS and resulted in the very fast response time, 0.29 mV/sec, 0.86 mV/sec at urea concentration of $50{\mu}g/ml,\; 500{\mu}g/ml$, respectively. And also in order to measure the uranyl ion, the uranyl ion selective sensing membrane with calix[6]arene derivative was used and its sensitivity was 25mV/concentration decade in the wide uranyl ion concentration range of $10^{-11}M\;to\;10^{-4}M$.