• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.698-706
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• 2004

$\beta$-Lactam antibiotics such as penicillin G, amoxicillin, and ampicillin were determined by a potentiometric biosensor system which exploited penicillinase immobilized on Immobilon cellulose nitrate membrane and a flat-bottomed pH electrode-as the biological component and transducer. The optimum reaction buffer for maximum sensitivity was found as 2 mM of sodium phosphate buffer (pH 7.2). The detection limit of the biosensor could be extended to 1 $\mu{M}$ of the analytes by increasing the enzyme loading for immobilization to 100 units/$m\ell$. The model samples spiked with each of the standard penicillins were measured for their biosensor responses and HPLC peak area, resulting in the relative responses of 82.1-103.5% and 79.5-106.1% for the biosensor method along with HPLC analysis, respectively. This result showed a good precision of the current biosensor method for screening the penicillin compounds.

• 한국식품과학회지
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• 제38권2호
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• pp.169-175
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• 2006

Citrate Iyase(CL)와 oxaloacetate decarboxylase(OD)를 고정화하여 효소 반응기를 제작하고 효소반응으로 생성된 carbonate ion을 ion selective electrode를 이용한 전위차법 FIA system으로 식품에함유된 구연산의 농도를 측정하였다. 최적 센서 시스템의 조건은 CL과 OD을 같이 고정화시키는 방법으로 CL과 OD의 양이 각각 10 units, 60 units, 담체량은 0.3 g, carrier buffer 1(0.1 MTris buffer)은 pH 7.5, carrier buffer의 유속은 12 mL/hr으로 결정되었다. 최적 상태에서의 표준검량곡선은 $10^{-1}\;M-10^{-3}\;M$ 범위에서 직선적인 상관관계$(r^2=0.9986)$를 나타내었다. 당류와 유기산류에 대한 센서 시스템에 대한 방해효과를 측정한 결과 당류는 거의 저해효과를 보이지 않았으며 유기산의 경우도 저해 효과가 5% 미만으로 이들 물질에 의해 감응도가 크게 방해받지 않은 것으로 나타났다. 구연산 바이오센서와 GC를 사용하여 식품 시료에 함유된 구연산의 농도를 분석한 결과 두 방법간에 유의적인 차이가 없는 것으로 나타나 본 연구에서 구축한 구연산 바이오센서 시스템은 식품에 함유된 구연산 분석시 신뢰성 있는 결과를 주는 것으로 보여졌다.

• BMB Reports
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• 제31권3호
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• pp.296-302
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• 1998

A potentiometric flow injection biosensor for the analysis of anti-cholinesterases (anti-ChEs), based on inhibition of enzyme activity, was developed. The sensor system consists of a reactor with acetylcholinesterase (AChE) immobilized on controlled pore glass and a detector with an $H^{+}-selective$ PVC-based membrane electrode. The principle of the analysis is based on the fact that the degree of inhibition of AChE by an anti-ChE is dependent on the concentration of the anti-ChE in contact with AChE. The sensor system was optimized by changing systematically the operating parameters of the sensor to evaluate the effect of the changes on sensor response to ACh. The optimized biosensor was applied to the analysis of paraoxon, an organophosphorus pesticide. Treatment of the inhibited enzyme with pyridine-2-aldoxime fully restored the enzyme activity allowing repeated use of the sensor.

• Preventive Nutrition and Food Science
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• 제4권1호
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• pp.79-83
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• 1999

A potentiometric biosensor employing a CO3-2 ion-selective electrode(ISE) and malic enzyme immobilization in al flow injection analysis (FIA) system was constructed. Analytical parameters were optimized for L-malate determination . The CO3-2 -ISE-FIA system was composed of a pump, an injector, a malic enzyme (EC1.1.1.40) reactor, a CO3-2 ion-selective electrode, a pH/mV meter and a recorder. Cofactor NADP was also injected with substrate for theenzyme reaction into the system. Optimized analytical parameters for L-malate determination in the CO3-2 ISE-FIA system were as follows ; flow rate, 14.5ml/hr ; sample injection volume, 100ul; enzyme loading in the reactor, 20 units ; length of the enzyme reactor , 7 cm ; tubing length form the enzyme reactor to the detector as a geometric factor in FIA, 15 cm . The response time for measuring the entire L-malate concentration range (10-2 ~10-5 mol/L ; 4 injections )was <15minutes . In this CO3-2 -ISE-FIA system, the potential differences due to th eformation of CO3-2 by the reaction of malic enzyme on L-malate were correlated to L-malate concentration in the range of 10-2 ~10-5mol/L ; the detection limit was 10-5 mol/L. This potentionmetric CO3-2 ISE--FIA system was found to be useful for L-malate measurement.

• BMB Reports
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• 제31권1호
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• pp.101-105
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• 1998

Immunoassay of botulinum toxin (BTX) B type was investigated using two typed of biosensors: light addressable potentiometric sensor (LAPS) and surface plasmon resonance (SPR) sensor. Urease-tagged and immuno-filtration capture method have been used for LAPS. Tag-free and direct binding real-time detection method have been used for SPR sensor. The detection limit of sandwich assay format with LAPS was 10 ng/ml, which was the lowest among methods tested. SPR has the advantage of being more convenient because tag-free direct binding assay can be used and reaction time was reduced, regardless of low sensitivity. This result shows that sandwich assay format with LAPS can be used as an alternative method of BTX mouse bioassay which is known as the most sensitive method for the detection of BTX.

• Preventive Nutrition and Food Science
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• 제3권1호
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• pp.36-42
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• 1998

Ion-selective eleltrodes(ISEs) are simple electrodechemical devices for the direct measurement of ions in the samples. A novel potentiometric biosensor for the determination of L-Malate or D-isocitrate has been developed by using CO2-3 -ISE-FIA system was composed of a pump, an injector, a malic enzyme or isocitric dehydrogenase enzyme reactor, a CO2-3 -ISE, a pH/mV meter, and an integrater. The various factors, such as buffer capacity types of plstericizer and polymer, were optimized for the CO2-3 selectivity. In this novel CO2-3 --ISE-FIA system, the potential difference due to the amount of CO2-3 produced from each enzyme reaction was proportional to the amount of L-malate or D-isocitrate.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2001년도 하계학술대회 논문집 C
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• pp.1454-1456
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• 2001

신장병 진단에 중요한 요소의 농도를 측정하기 위한 요소 센서를 반도체 위에 개발이 연구의 목적이다. 센서의 감도 측정은 선형 전위 주사법(Linear sweep voltammtry)을 이용하였다. 선형 전위 주사법은 가역적이든 비가역적이든 관계없이 cottrell 식에 의한 전류와 농도의 직선관계로부터 감도를 측정할 수 있는 장점이 있고 또한 저 농도에서 민감하게 반응한다. 따라서 기존 전위차 측정형 바이오 센서(Potentiometric biosensor)에서 규명할 수 없는 감도 문제를 선형 전위 주사법으로 규명하고자 하였다. 센서전극은 p-type 실리콘 웨이퍼 위에 전극을 제작했다. 그 위에 cyclic voltammetry 법을 사용하여 전도성 고분자를 전기 중합 하였고, 그위에 다시 chronoamperometry법을 사용하여 우레아제를 고정화 하여 작업전극으로 제작하였다. 센서의 감도는 phosphate buffer 용액(pH7.4)속에서 온도 35$^{\circ}C$를 유지하며 측정하였다.

• BMB Reports
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• 제30권5호
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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.