• Title/Summary/Keyword: biosensor activity

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A Dipstick-Type Electrochemical Immunosensor for The Detection of The Organophosphorus Insecticide Fenthion

  • Cho, Young-Ae;Cha, Geun-Sig;Lee, Yong-Tae;Lee, Hye-Sung
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.743-746
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    • 2005
  • A dipstick-type immunochemical biosensor for the detection of the organophosphorus insecticide fenthion was developed using a screen-printed electrode system as an amperometric transducer with polyclonal antibodies against fenthion as a bioreceptor. The assay of the biosensor involved competition between the pesticide in the sample and pesticide-glucose oxidase conjugate for binding to the antibody immobilized on the membrane. This was followed by measurement of the activity of the bound enzyme by the supply of the enzyme substrate (glucose) and amperometric determination of the enzyme reaction product ($H_2O_2$). The activity of the bound enzyme was inversely proportional to the concentration of pesticide. The optimized sensor system showed a linear response against the logarithm of the pesticide concentration ranging from $10^{-2}$ to $10^3\;{\mu}g/L$.

The Studies for the Malate Tissue Biosensor Using Malate Dehydrogenase(Decarboxylating) in the Bundle Sheath Cell of the Corn Leaf (옥수수잎의 유관속초세포내에 들어 있는 Malate Dehydrogenase(Decarboxylating)을 이용한 Malate 측정용 조직바이오센서에 관한 연구)

  • 김의락;노광수
    • KSBB Journal
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    • v.9 no.3
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    • pp.319-324
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    • 1994
  • A biosensor for the measurement of malate has been constructed by the sodium-alginate immobilized bundle sheath cell tissue of corn leaf containing malate dehydrogenase (decarboxylating) (EC 1. 1. 1. 40) on the CO2 gas-sensing electrode. The proposed tissue sensor had the linear in the range of malate concentration $5.5{\times}10^{-5}M∼2.5{\times}10^{-2}M$ with a slope of 53.5 mV/decade in 0.02M Tris-HCl buffer solution at optimum pH 8.0, and $25^{\circ}C$. A response time was 16∼18min. The present L-malate sensing tissue sensor is stable for more than one week. At pH 7.4, Km value was $0.6{\times}10^{-5}M$. The various kinds of salt did not effect the signal of malate tissue biosensor as the inhibitor. We can measure the malate by the CO2 electrode at the pH=8.0. Thus, the proposed tissue sensor will be useful for the measurement of malate.

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Immobilization of Recombinant Bacterial Biosensors: a Simple Approach for the On-Site Detection of Phenolic Compounds (재조합 박테리아 바이오센서의 고정화: 페놀계 화합물의 현장 검출을 위한 간단한 접근 방법)

  • Shin, Hae-Ja
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1323-1328
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    • 2011
  • We herein report the development of an agarose-gel-immobilized recombinant bacterial biosensor simple system for the field monitoring of phenolic compounds. Escherichia coli cells harboring the pLZCapR plasmid, which was previously designed to express the ${\beta}$-galactosidase reporter gene in the presence of phenolic compounds, were co-immobilized with a substrate [chlorophenol red ${\beta}$-galactopyranoside (CPRG) in agarose gel, and dispensed to the wells of a 96-well plate. Field samples were added to the wells and color development was monitored. In the presence of 5 ${\mu}M$ to 10 mM of phenol, the biosensor developed a red (representing hydrolysis of CPRG) color. Other phenolic compounds were also detected by this immobilized system, with the pattern resembling that previously reported for the corresponding non-immobilized biosensor. The immobilized cells showed optimum activity when the gel was simultaneously supplemented with 6% dimethyl formamide (DMF), 0.1% SDS and 10 mM $CaCl_2$. The immobilized biosensor described herein does not require the addition of a substrate or the use of unwieldy instruments or sample pretreatments that could complicate field studies.

A Recombinant Microbial Biosensor for Cadmium and Lead Detection (카드뮴 및 납 검출을 위한 재조합 미생물 바이오센서)

  • Shin, Hae Ja
    • Journal of Life Science
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    • v.26 no.5
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    • pp.503-508
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    • 2016
  • Biosensors have been used as first-step monitoring tools to detect on-site samples in a simple and cost-effective manner. Numerous recombinant microbial biosensors have been exploited for monitoring on-site toxic chemicals and biological signals. Herein, a recombinant microbial biosensor was constructed for monitoring cadmium. The cadmium responding cadC regulatory gene and it’s promoter from Staphylococcus aureus was amplified through PCR, fused with the lacZ gene, and transformed into Escherichia coli BL21 (DE3) cells. In the presence of cadmium, the biosensor cells express β-galactosidase showing red color development with chlorophenol red β-galactopyranoside (CPRG) as the enzymatic substrate. The biosensor cells showed the best β-galactosidase activity after 3 hr induction with cadmium at pH 5 and a detection range from 0.01 μM to 10 mM cadmium with a linearity from 0.01 to 0.1 μM cadmium (y = 0.98 x + 0.142, R2 = 0.98). Among the heavy metals, cadmium and lead showed good responses, tin and cobalt showed medium responses, and mercury and copper showed no responses. The biosensor cells showed good responses to several waste waters similar to buffer solution, all spiked with cadmium. The biosensor described herein could be applied for on-site cadmium monitoring in a simple and cost-effective manner without sample pretreatments.

QCM biosensor for the determination of haptoglobin (헵토글로빈 측정을 위한 수정미소저울 바이오센서 개발)

  • Kim, Sung-Il;Lee, Hyeon-Jeong;Park, Soon-Hyuk;Bhang, Hyun-Soo;Lee, Jin-Ah;Choi, Suk-Jung
    • Journal of Sensor Science and Technology
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    • v.16 no.2
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    • pp.132-141
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    • 2007
  • The purpose this research is to develop QCM (quartz crystal microbalance) biosensor for the determination of haptoglobin. Haptoglobin is an acute-phase protein with a hemoglobin-binding activity and has a potential to be used as a biomarker for infection or cancer. Haptoglobin level in milk has been used for the diagnosis of cow mastitis. In this study, anti-bovine haptoglobin antibody or bovine hemoglobin was chemically immobilized on the surface of the QCM, and the resulting sensor chips were tested for their response to samples containing haptoglobin at different concentrations. Concentration dependent frequency change was observed with both of the sensor chips. Especially, the sensor chip containing anti-bovine haptoglobin antibody showed sufficient sensitivity in the concentrations typically observed in the cows with mastitis.

Direct electrochemistry of hemoglobin at carbon electrode modified with lipid film and its application as a $H_{2}O_{2}$ sensor (Lipid Film에 수식된 헤모글로빈의 전기화학적 특성과 $H_{2}O_{2}$응답특성)

  • Lee, Dong-Yun;Park, Sang-Hyun;Choi, Yong-Sung;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2006.10a
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    • pp.93-94
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    • 2006
  • In this research, the enhancement of electron-transfer activity of hemoglobin (Hb) in dodecanoic acid film was investigated for the first time. This type of composite film was made on glassy carbon electrode by casting method. Cyclic voltammetric result of the modified electrode displays a well defined redox peaks which was attributed to the direct electrochemical response of Rb. Our results illustrate that Rb exchange electrons directly with electrode and exhibits the characteristics of peroxidase. When we apply this modified electrode as a biosensor, it gives excellent performances in the electrocatalytic reduction of hydrogen peroxide ($H_{2}O_{2}$). Through the optimal conditions, the proposed biosensor shows the linear range for H2O2 determination was from $1{\times}10^{-5}$ to $1.25{\times}10^{-4}mol/L$ with a detection limit of $1{\times}10^{-7}mol/L$. The biosensor retained more than 90% of the initial response after 14 days.

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A Study on Performance Improvement of Glucose Sensor Adopting a Catalyst Using New Cross Liker (새로운 가교제를 적용한 촉매를 이용한 글루코스 센서의 성능향상 연구)

  • Chung, Yongjin;Kwon, Yongchai
    • Korean Chemical Engineering Research
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    • v.53 no.6
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    • pp.802-807
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    • 2015
  • In this study, we synthesized a new biocatalyst consisting of glucose oxidase (GOx), polyethyleneimine (PEI) and carbon nanotube (CNT) with addition of terephthalaldehyde (TPA) (TPA/GOx/PEI/CNT) for fabrication of glucose sensor that shows improved sensing ability and stability compared with that using other biocatalysts. Main bonding of the new TPA/GOx/PEI/CNT catalyst is formed by Aldol condensation reaction of functional end groups between GOx/PEI and TPA. Such formed bonding structure promotes oxidation reaction of glucose. Catalytic activity of TPA/GOx/PEI/CNT is evaluated quantitatively by electrochemical measurements. As a result of that, large sensitivity value of $41{\mu}Acm^{-2}mM^{-1}$ is gained. Regarding biosensor stability of TPA/GOx/PEI/CNT catalyst, covalent bonding formed between GOx/PEI and TPA prevents GOx molecules from becoming leaching-out and contributes improvement in biosensor stability. With estimation of the biosensor stability, it is found that the TPA/GOx/PEI/CNT catalyst keeps 94.6% of its initial activity even after three weeks.

A Study on the Electrochemical Characteristics of Biosensor with HRP Enzyme Immobilized on SPCE (SPCE에 HRP 효소가 고정화된 바이오센서의 전기화학적 특성에 관한 연구)

  • Han, Kyoung Ho;Lee, Dae Hyun;Yoon, Do Young;Choi, Sangil
    • Journal of the Korean Electrochemical Society
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    • v.23 no.3
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    • pp.73-80
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    • 2020
  • Fenton oxidation method using hydrogen peroxide is an eco-friendly oxidation method used in water treatment and soil restoration. When removing pollutants by this method, it is quite important to properly regulate the concentration of hydrogen peroxide according to the concentration of the contaminants. In this study, electrochemical biosensors using HRP (horseradish peroxidase) enzymes were manufactured and studies were conducted on the activity of enzymes and the detection characteristics of hydrogen peroxide. HRP were electro deposited with chitosan and AuNP on the working electrode surface of the SPCE (Screen Printed Carbon Electrode). Then, the fixation of enzymes was confirmed using the cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The activity of HRP enzymes was also identified from chronoamperometry (CA) and UV spectroscopy. After immersing the biosensor in PBS solution the current generated from electrodes by titrating hydrogen peroxide was measured from CA analysis. The generated current increased linearly for the concentration of hydrogen peroxide, and a calibration curve was derived that could predict the concentration of hydrogen peroxide from the current.

Studies on Lytic, Tailed Bacillus cereus-specific Phage for Use in a Ferromagnetoelastic Biosensor as a Novel Recognition Element

  • Choi, In Young;Park, Joo Hyeon;Gwak, Kyoung Min;Kim, Kwang-Pyo;Oh, Jun-Hyun;Park, Mi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.87-94
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    • 2018
  • This study investigated the feasibility of the lytic, tailed Bacillus cereus-specific phage for use in a ferromagnetoelastic (FME) biosensor as a novel recognition element. The phage was immobilized at various concentrations through either direct adsorption or a combination of 11-mercapto-1-undecanoic acid (11-MUA) and [N-(3-dimethylaminopropyl)-N'-carbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS)]. The effects of time and temperature on its lytic properties were investigated through the exposure of B. cereus (4 and 8 logCFU/ml) to the phage (8 logPFU/ml) for various incubation periods at $22^{\circ}C$ and at various temperatures for 30 and 60 min. As the phage concentration increased, both immobilization methods also significantly increased the phage density (p < 0.05). SEM images confirmed that the phage density on the FME platform corresponded to the increased phage concentration. As the combination of 11-MUA and EDC/NHS enhanced the phage density and orientation by up to 4.3-fold, it was selected for use. When various incubation was conducted, no significant differences were observed in the survival rate of B. cereus within 30 min, which was in contrast to the significant decreases observed at 45 and 60 min (p < 0.05). In addition, temperature exerted no significant effects on the survival rate across the entire temperature range. This study demonstrated the feasibility of the lytic, tailed B. cereus-specific phage as a novel recognition element for use in an FME biosensor. Thus, the phage could be placed on the surface of foods for at least 30 min without any significant loss of B. cereus, as a result of the inherent lytic activity of the B. cereus-specific phage as a novel recognition element.