• Title/Summary/Keyword: enzyme sensor

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Signal Analysis of Optical Biosensor to Detect Peroxide Using Electrically Controlled Release System (전기적 방출 조절 시스템을 이용한 광 페록사이드센서의 개발)

  • Min, Jun-Hong;Lim, In-Hee;Kim, Hyo-Han;Lee, Sang-Beak;Choi, Jeong-Woo;Lee, Won-Hong
    • Journal of Sensor Science and Technology
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    • v.6 no.1
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    • pp.35-42
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    • 1997
  • The optical biosensor using the electrically controlled release of reactive reagent is developed for the detection of peroxide. Rapid degradation of polymer complex of PEOx and PMAA occurs as the applied current increases and thus released amount of HPA increases. The degradation velocity of polymer and the amount of HPA released are linearly proportional to the applied current. Peroxide is reacted with the released reagent by peroxidase and then the product, a fluorescent dimer DBDA, is formed. The monochromic light from light source (150W Xe arc ramp) excites the DBDA and the excited light is transmitted through an optical fiber to be detected by a photodiode array. The change of fluorescence intensity is related to the change of peroxide concentration. The peroxidase is entrapped in Ca-alginate get on the inner surface. The biosensor has the linear signal range of 0.025mM-10.mM peroxide. By applying the step function of peroxide, reproducibility of biosensor has been investigated. The mathematical model is constructed by the combination of enzyme kinetics with reactor flow model. Good agreement is obtained between the experimental result and model prediction in the sensor signal.

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An ELISA-on-a-Chip Biosensor System for Early Screening of Listeria monocytogenes in Contaminated Food Products

  • Seo, Sung-Min;Cho, Il-Hoon;Kim, Joo-Ho;Jeon, Jin-Woo;Oh, Eun-Gyoung;Yu, Hong-Sik;Shin, Soon-Bum;Lee, Hee-Jung;Paek, Se-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2993-2998
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    • 2009
  • An enzyme-linked immunosorbent assay (ELISA)-on-a-chip (EOC) biosensor combined with cell concentration technology based on immuno-magnetic separation (IMS) was investigated for use as a potential tool for early screening of Listeria monocytogenes (L. monocytogenes) in food products. The target analyte is a well-known pathogenic foodborne microorganism and outbreaks of the food poisoning typically occur due to contamination of normal food products. Thus, the aim of this study was to develop a rapid and reliable sensor that could be utilized on a daily basis to test food products for the presence of this pathogenic microorganism. The sensor was optimized to provide a high detection capability (e.g., 5.9 ${\times}\;10^3$ cells/mL) and, to eventually minimize cultivation time. The cell density was condensed using IMS prior to analysis. Since the concentration rate of IMS was greater than 100-fold, this combination resulted in a detection limit of 54 cells/mL. The EOC-IMS coupled analytical system was then applied to a real sample test of fish intestines. The system was able to detect L. monocytogenes at a concentration of 2.4 CFU/g after pre-enrichment for 6 h from the onset of cell cultivation. This may allow us to monitor the target analyte at a concentration less than 1 CFU/g within a 9 h-cultivation provided a doubling time of 40 min is typically maintained. Based on this estimation, the EOC-IMS system can screen and detect the presence of this microorganism in food products almost within working hours.

Enzyme-Free Glucose Sensing with Polyaniline-Decorated Flexible CNT Fiber Electrode (Polyaniline을 이용한 CNT fiber 유연 전극 기반의 비효소적 글루코스 검출)

  • Song, Min-Jung
    • Korean Chemical Engineering Research
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    • v.60 no.1
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    • pp.1-6
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    • 2022
  • As the demand for wearable devices increases, many studies have been studied on the development of flexible electrode materials recently. In particular, the development of high-performance flexible electrode materials is very important for wearable sensors for healthcare because it is necessary to continuously monitor and accurately detect body information such as body temperature, heart rate, blood glucose, and oxygen concentration in real time. In this study, we fabricated the nonenzymatic glucose sensor based on polyaniline/carbon nanotube fiber (PANI/CNT fiber) electrode. PANI layer was synthesized on the flexible CNT fiber electrode through electrochemical polymerization process in order to improve the performance of a flexible CNT fiber based electrode material. Surface morphology of the PANI/CNT fiber electrode was observed by scanning electron microscopy. And its electrochemical characteristics were investigated by chronoamperometry, cyclic voltammetry, electrochemical impedance spectroscopy. Compared to bare CNT fiber electrode, this PANI/CNT fiber electrode exhibited small electron transfer resistance, low peak separation potential and large surface area, resulting in enhanced sensing properties for glucose such as wide linear range (0.024~0.39 and 1.56~50 mM), high sensitivity (52.91 and 2.24 ㎂/mM·cm2), low detection limit (2 μM) and good selectivity. Therefore, it is expected that it will be possible to develop high performance CNT fiber based flexible electrode materials using various nanomaterials.

Electrochemical Immobilization of Osmium Complex onto the Carbon Nano-Tube Electrodes and its Application for Glucose sensor (전기화학적인 방법을 이용한 탄소나노튜브 전극상의 오스뮴 착물의 고정화 및 혈당센서에 관한 응용)

  • Choi, Young-Bong;Jeon, Won-Yong;Kim, Hyug-Han
    • Journal of the Korean Electrochemical Society
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    • v.13 no.1
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    • pp.50-56
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    • 2010
  • The multi-wall carbon nano-tube composite mixed with carbon paste electrode presented more sensitive and selective amperometric signals in the oxidation of glucose than general screen-printed carbon electrodes(SPCEs). Redox mediators to transport electrodes from enzyme to electrodes are very important part in the biosensor. A novel osmium redox complex was synthesized by the coordinating pyridine group containing primary amines which were electrochemically immobilized onto the MWCNT-SPCEs surface. Electrochemical studies of osmium complexes were investigated by cyclic voltammetry, chronoamperometry. The surface coverage of osmium complexes on the modified carbon nano-tube electrodes were significantly increased at 100 time (${\tau}_0=2.0\;{\times}\;10^{-9}\;mole/cm^2$) compared to that of the unmodified carbon electrodes. It's practical application of the glucose biosensor demonstrated that it shows good linear response to the glucose concentration in the range of 0-10 mM.

Electrochemical Immunosensor Based on the ZnO Nanorods Inside PDMS Channel for H7N9 Influenza Virus Detection (PDMS 채널 내부에 성장된 산화아연 나노막대를 이용한 H7N9 인플루엔자 바이러스 전기화학 면역센서)

  • Han, Ji-Hoon;Lee, Dongyoung;Pak, James Jungho
    • Journal of Sensor Science and Technology
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    • v.23 no.4
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    • pp.278-283
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    • 2014
  • In this study, we propose an immunosensor using zinc oxide nanorods (NRs) inside PDMS channel for detecting the influenza A virus subtype H7N9. ZnO with high isoelectric point (IEP, ~9.5) makes it suitable for immobilizing proteins with low IEP. In this proposed H7N9 immunosensor structure ZnO NRs were grown on the PDMS channel inner surface to immobilize H7N9 capture antibody. A sandwich enzyme-linked immunosorbent assay (ELISA) method with was used 3,3',5,5' tetramethylbenzidine (TMB) for detecting H7N9 influenza virus. The immunosensor was evaluated by amperometry at various H7N9 influenza antigen concentrations (1 pg/ml - 1 ng/ml). The redox peak voltage and current were measured by amperometry with ZnO NWs and without ZnO NWs inside PDMS channel. The measurement results of the H7N9 immunosensor showed that oxidation peak current of TMB at 0.25 V logarithmically increased from 2.3 to 3.8 uA as the H7N9 influenza antigen concentration changed from 1 pg/ml to 1 ng/ml. And then we demonstrated that ZnO NRs inside PDMS channel can improve the sensitivity of immunosensor to compare non-ZnO NRs inside PDMS channel.

Dopamine determination using a biosensor based on multiwall carbon nanotubes paste and burley tobacco-peroxidase (담배 잎-peroxidase와 다중벽 탄소 나노튜브를 이용한 dopamine의 정량)

  • Kwon, Hyoshik;Jeon, Byong-Suk;Pak, Yongnam
    • Analytical Science and Technology
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    • v.28 no.2
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    • pp.98-105
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    • 2015
  • The development of an enzymatic biosensor for dopamine determination based on multiwall carbon nanotubes (MWCNTs) and peroxidase obtained from the crude extract of burley tobacco (Nicotiana tabacum L.) was proposed. Peroxidase catalyzes the oxidation of dopamine to dopamine quinone. The influence on the response of analytical parameters of biosensors such as enzyme concentration, dopamine concentration, pH, and phosphate buffer solution concentration were investigated. The analytical parameters obtained, including sensitivity, linearity, and stability, were investigated. The proposed method for dopamine determination presented good selectivity even in the presence of uric acid and ascorbic acid. The sensor presented a higher response for dopamine in 0.010 M phosphate buffer at pH 6.50, with an applied potential of -0.15 V. The detection limit of the electrode was 2.7×10−6 M (S/N = 3) and the relative standard deviation of the measurements, which were repeated 10 times using 5.0×10−2 M dopamine, was 1.3%.

Identification of another calmodulin-binding domain at the C-terminal region of AtCBP63

  • Kim, Sun-Ho;Kang, Yun-Hwan;Han, Hay-Ju;Bae, Dong-Won;Kim, Min-Chul;Lim, Chae-Oh;Chung, Woo-Sik
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.53-58
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    • 2009
  • Calcium signals can be transduced by binding calmodulin (CaM), a $Ca^{2+}$ sensor in eukaryotes, is known to be involved in the regulation of diverse cellular functions. We isolated a CaM-binding protein 63 kD (AtCBP63) from the pathogen-treated Arabidopsis cDNA expression library. Recently, AtCBP63 was identified as a CaM bining protein. The CaM binding domain of AtCBP63 was reported to be located in its N-terminal region, In this study, however, we showed that ACaM2 could specifically bind to second CaM-binding domain (CaMBD) of AtCBP63 at the C-terminal region. The specific binding of CaM to CaM binding domain was confirmed by a gel mobility shift assay, a split ubiquitin assay, site-directed mutagenesis, and a competition assay using a $Ca^{2+}$/CaM-dependent enzyme. The gene expression of AtCBP63 was induced by pathogens and pathogens related second messengers. This result suggests that a CaM binding protein, AtCBP63, may play role in pathogen defense signaling pathway.

Development of HRP-modified Carbon Composite Biosensor and Electrochemical Analysis of H2O2 (Horseradish peroxidase가 변성된 탄소복합 바이오센서 개발 및 전기화학적 H2O2분석)

  • Park, Deog-Su
    • Journal of the Korean Chemical Society
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    • v.56 no.5
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    • pp.571-576
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    • 2012
  • A sol-gel derived carbon composite electrodes (CCEs) were fabricated by mixing horseradish peroxidase (HRP), sol of tetraethoxysilane (TESO), and graphite powder. The HRP solution was added to the sol solution of TEOS, and then graphite powder was added to this mixture. The resulting carbon ceramic network effectively encapsulated HRP and shows a catalytic reduction starting at -0.2 V for $H_2O_2$. The optimum conditions for $H_2O_2$determination have been characterized with respect to the enzyme loading ratio and pH. The linear range and detection limit of $H_2O_2$ detection were from 0.2 mM to 2.2 mM and 0.035 mM, respectively. The common electroactive interferences such as ascorbic acid, acetaminophene, and uric acid were not affected upon the response to $H_2O_2$ at the HRP biosensor due to low detection potential.

An Aptamer-Based Electrochemical Sensor That Can Distinguish Influenza Virus Subtype H1 from H5

  • Lee, Jin-Moo;Kim, JunWon;Ryu, Ilhwan;Woo, Hye-Min;Lee, Tae Gyun;Jung, Woong;Yim, Sanggyu;Jeong, Yong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.27 no.11
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    • pp.2037-2043
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    • 2017
  • The surface protein hemagglutinin (HA) mediates the attachment of influenza virus to host cells containing sialic acid and thus facilitates viral infection. Therefore, HA is considered as a good target for the development of diagnostic tools for influenza virus. Previously, we reported the isolation of single-stranded aptamers that can distinguish influenza subtype H1 from H5. In this study, we describe a method for the selective electrical detection of H1 using the isolated aptamer as a molecular probe. After immobilization of the aptamer on Si wafer, enzyme-linked immunosorbent assay (ELISA) and field emission scanning electron microscopy (FE-SEM) showed that the immobilized aptamer bound specifically to the H1 subtype but not to the H5 subtype. Assessment by cyclic voltammetry (CV) also demonstrated that the immobilized aptamer on the indium thin oxide-coated surface was specifically bound to the H1 subtype only, which was consistent with the ELISA and FE-SEM results. Further measurement of CV using various amounts of H1 subtype provided the detection limit of the immobilized aptamer, which showed that a nanomolar scale of target protein was sufficient to produce the signal. These results indicated that the selected aptamer can be an effective probe for distinguishing the subtypes of influenza viruses by monitoring current changes.

A Biosensor for the Rapid Detection of the Fungicide Iprovalicarb Residuess (살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 바이오센서)

  • Cho, Han-Keun;Kim, Woon-Ho;Kyung, Kee-Sung;Lee, Eun-Young
    • Journal of Biosystems Engineering
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    • v.32 no.6
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    • pp.440-447
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    • 2007
  • In this study, a biosensor was developed using an enzyme-linked immunosorbent assay (ELISA) to rapidly measure the fungicide iprovalicarb residues in agricultural products. The biosensor was designed to include micro-pumps and solenoid valves for fluid transport, a spectrophotometer cuvet as a reaction chamber, a photodiode with a light-emitting diode for optical density measurement, and a control microcomputer to implement assay. The rate of change in optical density of the cuvet was read as final signal output. Micro-pumps were evaluated to investigate their delivery capability, the highest values of the error and the coefficient of variation were 4.3% and 4.6% respectively. As the incubation period was reduced from 15 minutes to 11 minutes to shorten the total processing time, the sensor sensitivity was decreased as the antibody dilution ratio was reduced to a half. The maximum usable period of the coated cuvet was found to be two days with 1% error limit. To predict the concentration of the iprovalicarb residue in agricultural products, a linear calibration model was obtained with r-square values of 0.992 for potato and 0.985 for onion. In validation test for the samples of potatoes and onions against the high performance liquid chromatography, very high correlation values were obtained as 0.996 and 0.993 respectively. Using the cuvet immobilized with antigen, it took 21-minutes for the biosensor to complete the measuring process of the iprovalicarb residues.