• Journal of Microbiology and Biotechnology
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• 제33권2호
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• pp.228-234
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• 2023

In this study, the performance of a gold biosensor combined with light microscope imaging system (GB-LMIS) was comparatively evaluated against enzyme-linked immunosorbent assay (ELISA) for detecting Salmonella under simulated chilling condition. The optimum concentration of antiSalmonella polyclonal antibodies (pAbs) was determined to be 12.5 and 100 ㎍/ml for ELISA and GBLMIS, respectively. GB-LMIS exhibited a sufficient and competitive specificity toward three tested Salmonella among only. To mimic a real-world situation, chicken was inoculated with Salmonella cocktail and stored under chilling condition for 48 h. The overall growth of Salmonella under chilling condition was significantly lower than that under non-exposure to the chilling condition (p < 0.05). No significant differences in bacterial growth were observed between brain heart infusion and brilliant green broth during the enrichment period (p > 0.05). Finally, both GB-LMIS and ELISA were employed to detect Salmonella at every 2-h interval. GB-LMIS detected Salmonella with a competitive specificity by the direct observation of bacteria on the sensor using a charge-coupled device camera within a detection time of ~2.5 h. GB-LMIS is a feasible, novel, and rapid method for detecting Salmonella in poultry facilities.

• 한국식품과학회지
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• 제30권1호
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• pp.22-34
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• 1998

고정화효소와 산소전극 시스템을 이용한 효소센서를 제작하여 식품 중의 당, 유기산, 알코올 성분을 동시 측정 하였다. 효소가 기질과 반응하여 소비한 산소의 변화량이 전압차이로 나타나므로 시간당 전압 감소량이 최대인 값으로부터 각 성분의 농도를 측정하였으며, 이때 1분내에 최대기울기를 구할 수 있어 신속한 측정이 가능하였다. 효소의 고정화 지지체로는 nylon cloth를 사용하였고, asymmetrical coupling 방법에 의하여 기질 작용 순으로 위치하도록 효소를 고정화하였다. 한 개의 양극과 6개의 음극으로 제작된 multiple cathode system으로 포도당, 젖산, 에탄올 성분을 동시 측정할 수 있는 효소 센서를 제작하였다. 위의 센서 제작을 위하여 mutarotase과 glucose oxidase/lactate oxidase/alcohol oxidase와 catalase가 각기 사용되었다. 이들 효소센서의 최적조건은 $pH\;7.0,\;40^{\circ}C$의 0.1 M 인산완충용액이었으며 각 효소 센서의 방해물질을 알아 보기 위하여 여러 가지 당과 각종 유기산, 알콜류에 대한 효소 감응도를 살펴 본 결과 포도당 센서에서 유기산의 영향을 제외하고는 10% 내외였다. 따라서 포도당과 유기산을 동시 측정하기 위하여 포도당/젖산의 영향을 고려한 적절한 보정관계식을 도입하여 순수한 유리당과 유기산의 값을 측정할 수 있었다. 제작된 효소센서의 검증을 위하여 분광광도법. HPLC, GC를 이용한 결과, 분석방법간에 높은 상관관계를 보여 주었다. 아울러 각 효소센서의 안정성을 살펴본 결과 알코올 센서를 제외하고는 30일 이후에도 80%이상 효소감응도가 유지되었다.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.594-599
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• 2007

It is well known that the Escherichia coli inner membrane-bound protease DegS is a periplasmic stress sensor for unfolded outer membrane proteins (OMPs). Previous studies have also shown that the outer membrane protease OmpT activates plasminogen in vitro and this may be exploited by bacteria in the course of pathogenesis. However, there has been no research on the plasminogen activation ability of the important periplasmic protein DegS. Accordingly, in this study, the whole-length and truncated degS genes were separately overexpressed in Escherichia coli, the recombinant proteins purified by affinity chromatography, and their plasminogen activator role tested in vitro. The results suggested that the whole-length DegS was able to activate plasminogen on a plasma plate. The truncated form of DegS (residues 80-345), designated ${\Delta}DegS$, also acted as a plasminogen activator, as confirmed by different assays. The serine protease property of ${\Delta}DegS$ was verified based on the complete inhibition of its enzyme activity by PMSF (phenylmethanesulfonyl fluoride). Therefore, the present results indicate that DegS is a plasminogen activator in vitro.

• 센서학회지
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• 제20권1호
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• pp.35-39
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• 2011

A disposable electrochemical immunosensor system has been developed for the detection of herbicide in aqueous samples. Disposable screen printed carbon electrodes(SPCE) were used as basic electrodes and an enzyme, horseradish peroxidase (HRP), and anti-herbicide antibodies was immobilised on to the working electrode of SPCE by using avidin-biotin coupling reactions. An herbicide-glucose oxidase conjugates have been used for the competitive immunoreaction with sample herbicides. The enzymatic reaction between the conjugated glucose oxidase and glucose added generates hydrogen peroxide, which was reduced by the peroxidase immobilised. The latter process caused an electrical current change, due to direct re-reduction of peroxidase by a direct electron transfer mechanism, which was measured to determine the herbicides in the sample. The optimal operational condition was found to be: $20\;{\mu}gl-1$ deglycosylated avidin loading to the working electrode and working potential +50 mV vs. Ag/AgCl. The total assay time was 15 min after sample addition. The detection limits for herbicides, atrazine and simazine, were found to be 3 ppb and 10 ppb, respectively.

• Journal of Electrochemical Science and Technology
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• 제2권3호
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• pp.163-167
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• 2011

Nicotinamide adenine dinucleotide, $NAD^+$, and its reduced form, NADH, play important roles as coenzymes in many enzymatic reactions. Electrochemical methods for $NAD^+$ or NADH detection or generation are drawn attention because it can provide the simple and low cost platform with fairly good sensitivity. In this study, the polysiloxane viologen polymer/diaphorase/hydrophilic polyurethane (PSV/DI/HPU) modified electrodes were simply prepared and demonstrated for bio-electrocatalytic $NAD^+$ sensors. The electrodes were co-immobilized with diaphorase and polysiloxane viologen polymer as an electron mediator followed by the overcoating with HPU membrane. The mixture of the enzyme and the electron mediator was well stabilized within HPU membrane and exhibited good reversibility and stability. The sensitivity was 0.2 $nA{\cdot}{\mu}M^{-1}$ and the detection limit was 28 ${\mu}M$ with a response time of 50 s ($t_{90%}$). The capability for NADH sensor was also observed on the PSV/DI/HPU electrode.

• 전기화학회지
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• 제25권1호
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• pp.13-21
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• 2022

There is an increasing interest in monitoring of specific biomarker for determining progression of a disease or efficacy of a treatment. Conventional method for quantification of specific biomarkers as enzyme linked immunosorbent assay (ELISA) has high material costs, long incubation periods, requires large volume of samples and involves special instruments, which necessitates clinical samples to be sent to a lab. This paper reports on the development of an electrochemical biosensor to measure total immunoglobulin E (IgE), a marker of asthma disease that varies with age, gender, and disease in concentrations from 0.3-1000 ng/mL with consuming 20 µL volume of whole blood sample. The sensor provides rapid, accurate, easy, point-of-care measurement of IgE, also, sequential monitoring of total IgE with ovalbumin (OVA) induced mice is another application of sensor. Taken together, these results provide an alternative way for detection of biomarkers in whole blood with low volumes and long-term ex-vivo assessments for understanding the progression of a disease.

• 전기화학회지
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• 제17권3호
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• pp.164-171
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• 2014

무효소 혈당센서는 높은 선택성과 민감성을 가지고 저비용으로 체내 혈당(glucose)을 검출할 수차세대 기술이다. 현재 시판되고 있는 혈당센서는 당을 산화시켜주는 당산화효소와 전극과 효소사이에 전자 전달을 원활하게 해주는 산화/환원 매개체를 이용하여 효소센서로 제작된다. 그러나 이러한 효소센서는 pH, 온도, 습도, 화학적 독성물질 등에 영향을 많이 받아 안정성이 떨어지고, 제작에 비용이 많이 드는 단점을 가지고 있다. 본 논문은 위와 같은 단점을 해결하고자 환원제인 당에 의하여 환원되는 니켈 나노입자를 전기화학적 흡착방법을 이용하여 산화 인듐 주석 전극 (ITO)에 고정시켰다. 고정된 니켈 나노입자는 전극의 표면적을 넓혀 신호를 증폭시키는 효과를 가지고 있으며, 당에 의하여 계속적으로 니켈이 환원됨에 따라 전극 반응에서는 촉매산화전류 반응으로 나타낸다. 당의 농도에 따라서 선형적으로 감응 할 수 있는 최적 조건의 니켈 나노입자를 이용하여 혈당센서를 제작하였다. 또한 체내에 존재하는 방해 인자인 아스코브산의 간섭을 억제하기 위해 음이온 고분자의 표면처리를 통하여 상대적으로 당에 선택적으로 감응하도록 하였다. 제작된 전극을 통하여 당 농도 별 산화 촉매 전류를 순환 전압 전류 법으로 측정한 결과 650 mV (vs. Ag/AgCl)에서 최대 전기적 신호가 발생되었으며, 포도당 0~6.15 mM 의 농도범위에서 전기적 신호가 선형 증가함을 확인할 수 있었다.

• 센서학회지
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• 제13권5호
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• pp.356-362
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• 2004

In this study, we developed the measuring system based on light addressable potentiometric sensor for the quantitative analysis of penicillin that is very important element in medicine and pharmacy, clinic. It is investigated the response characteristics by enzyme reaction with penicillinase. First, the surface pre-treatment process of the $Si_{3}N_{4}$ was established. The coupling agent was made using self assembled monolayer method and it was confirmed the immobilization process by AFM. Also, as the measuring system, potentiostat, signal processing part etc. was made by Lab VIEW software, it was reduced detecting time as well as simplifying the system. Fabricated device was shown excellent pH response characteristics, 57 mV/ pH in the range of pH $2{\sim}11$. The response characteristics was 60 mV/decade in the range of $0.1{\sim}10{\;}mM$.

• 분석과학
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• 제7권2호
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• pp.149-153
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• 1994

Squash-조직을 graphite rod disk 전극에 고정하여 메탄올 용매 속에서 L-ascorbic acid(AA)를 정량할 수 있는 전류법 바이오센서를 만들었다. 전극의 검출한계는 $2{\times}10^{-6}M$이었다. 순수한 ascorbate oxidase(AO)를 고정시켜 만든 전극에 비해 식물 조직에 들어 있는 효소로 만든 이 전극은 생촉매의 활성도가 높고 안정성이 좋았으며(1주간) 대단히 값싸게 만들 수 있었다.

• Archives of Pharmacal Research
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• 제20권1호
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• pp.68-71
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• 1997

Penicillin sensor was prepared by immobilizing penicillinase (Pcase) on $H^{+}$-selective carboxylated poly (vinyl chloride) (PVC-COOH) membrane or cellulose filter membrane. The immobilization techniques are as follows. Pcase was immobilized with GTH on $H^{+}$-selective PVC-COOH membrane or some amount of BSA was dropped on that membrane. Another method to make immobilization is to mix type I Pcase with GTH and drop on a cellulose filter membrane. According to immobilization techniques, there were some differences in response properties of enzyme electrodes, however, all electrodes responded to Pcase-resistant penicillin derivatives. Pcase immobilized on cellulose filter membrane with $H^{+}$-selective PVC membrane eletrode was more stable and more sensitive to penicillinase-resistant penicillin derivatives than any other immobilization techniques.