• Journal of Biosystems Engineering
• /
• 제28권2호
• /
• pp.167-172
• /
• 2003

This study was performed to fabricate a batch-type SPR biosensing system using a thiolated E. coli antibody coupling, and to explore the feasibility of real-time detection of E. coii in a stagnant sample solution. In advance. “O” and “K” antigenic serotype E. coli antibodies were thiolated with sulfo-LC-SPDP and dithiothreitol, and immobilized by chemisorption in the gold surface of compact SPR sensors. When the SPR biosensor immobilized with E. coli antibody monitored a E. coli solution, it took 3 to 5 min to stabilize. The SPR biosensing system developed in this study was able to detect E. coli in the range above 10$^4$ CFU/mL at the 0.05 significant level. Also, the SPR biosensor had possibility to significantly detect E. coli in the range of 10$^2$ to 10$^4$ CFU/mL in E. coli solutions. Meanwhile, when the SPR biosensor immobilized with 5. coli antibody was cleaned with NaOH solutions, its ability to detect E. coli largely decreased due to wash-out of the immobilized antibody. In order to reuse the SPR sensor, it should be antibody-immobilized newly.

• Journal of Biosystems Engineering
• /
• 제35권2호
• /
• pp.116-123
• /
• 2010

Rapid detection of foodborne pathogens has been a major challenge for the food industry. Salmonella contamination is well known in all foods including pasteurised milk. The possibility of specific detection of Salmonella Enteritidis by surface plasmon resonance (SPR) biosensor was explored using a commercially available portable SPR sensor. Self assembly technique was adopted to immobilize anti-Salmonella antibodies on the gold sensing surface of the SPR sensor. The concentration of polyclonal antibody for use in the SPR biosensor was chosen to 1.0 mg/mL. Experiments were conducted at near real-time with results obtained for one SPR biosensor assay within 1 hour. The limit of detection for Salmonella Enteritidis was determined to be $10^6$ CFU/mL in both PBS buffer and milk samples. The assay sensitivity was not significantly affected by milk matrix. Our results showed that it would be possible for employing the SPR biosensor to detect Salmonella Enteritidis in near real-time.

• Applied Science and Convergence Technology
• /
• 제25권3호
• /
• pp.61-65
• /
• 2016

Here, the rapid detection of Salmonella typhimurium by a portable surface plasmon resonance (SPR) biosensor in which the beam from a diode laser is modulated by a rotating mirror is reported. Using this system, immunoassay based on lipopolysaccharides (LPS)-specific monoclonal anti-Salmonella antibody was performed. For the purpose of orientation-controlled immobilization of antibodies on the SPR chip surface, the cysteine-mediated immobilization method, which is based on interaction between a gold surface and a thiol group (-SH) of cysteine, was adopted. As a result, using the portable SPR-based immunoassay, we detected S. typhimurium in the range from 10^7 CFU/mL to 10^9 CFU/mL within 1 hour. The results indicate that the portable SPR system could be potentially applied for general laboratory detection as well as on-site monitoring of foodborne, clinical, and environmental agents of interest.

• Journal of Biosystems Engineering
• /
• 제34권1호
• /
• pp.50-56
• /
• 2009

Surface plasmon resonance (SPR) biosensor has been used to detect many biochemical reactions, because this label-free sensor has high sensitivity and rapid response. The reactions are monitored by refractive index changes of the SPR biosensor. Iprovalicarb is protective, curative, and eradicative systemic fungicide introduced by Bayer AG in 1999. It has potential far control of downy mildew infesting onion, cucumber, grape and melon, late blight infesting tomato and potato, and anthracnose infesting watermelon and pepper. It is strictly limited to the maximum residue limit. In this study, the applicability of a portable SPR biosensor (Spreeta, Texas instrument, TX, USA) to detect the iprovalicarb residue was examined. The sensor chip was adopted to detect the reaction of iprovalicarb to immobilized iprovalicarb-antibody. The binding of the iprovalicarb onto the biosensor surface was measured by change of the refractive index (RI). Characteristics of the sensor chip including specificity, sensitivity, stability, and reusability were analyzed. In calibration test for seven levels of iprovalicarb concentration (0.32 to 5,000 mg/L) with three replications, a Sigmoidal model with Hill function was obtained between relative RI value and the iprovalicarb concentration with R-square of 0.998. It took 30 minutes to complete a set of detecting assay with the SPR biosensor.

• Food Science and Biotechnology
• /
• 제17권5호
• /
• pp.1038-1046
• /
• 2008

This study was carried out to develop a small-sized biosensor based on surface plasmon resonance (SPR) for the rapid identification of insecticide residues for food safety. The SPR biosensor module consists of a single 770 nm-light emitting diodes (LED) light source, several optical lenses for transferring light, a hemisphere sensor chip, photo detector, A/D converter, power source, and software for signal processing using a computer. Except for the computer, the size and weight of the sensor module are 150 (L)$\times$70 (W)$\times$120 (H) mm and 828 g, respectively. Validation and application procedures were designed to assess refractive index analysis, affinity properties, sensitivity, linearity, limits of detection, and robustness which includes an analysis of baseline stability and reproducibility of ligand immobilization using carbamate (carbofuran and carbaryl) and organophosphate (cadusafos, ethoprofos, and chlorpyrifos) insecticide residues. With direct binding analysis, insecticide residues were detected at less than the minimum 0.01 ppm and analyzed in less than 100 sec with a good linear relationship. Based on these results, we find that the binding interaction with active target groups in enzymes using the miniaturized SPR biosensor could detect low concentrations which satisfy the maximum residue limits for pesticide tolerance in Korea, Japan, and the USA.

• Journal of the Optical Society of Korea
• /
• 제13권3호
• /
• pp.392-397
• /
• 2009

We designed a wavelength interrogation-based surface plasmon resonance (SPR) biosensor to detect avian influenza DNA (AI-DNA). Hybridization reactions between target AI-DNA probes and capture probes immobilized on a gold surface were monitored quantitatively by measuring the resonance wavelength in the visible waveband. The experimental results were consistent with numerical calculations. Although the SPR detection technique does not require the DNA to be labeled, we also evaluated fluorescently-labeled targets to verify the hybridization behavior of the AI-DNA. Changes in resonance were found to be linearly proportional to the amount of bound analyte. A wavelength interrogation-type SPR biosensor can be used for rapid measurement and high-throughput detection of highly pathogenic AI viruses.

• 한국미생물·생명공학회지
• /
• 제49권4호
• /
• pp.510-518
• /
• 2021

Bacteriophages are considered excellent sensing elements for platforms detecting bacteria. However, their lytic cycle has restricted their efficacy. Here, we used the minor coat protein pIII domain (N1N2) of phage CTXφ to construct a novel surface plasmon resonance (SPR) biosensor that could detect Vibrio cholerae. N1N2 harboring the domains required for phage adsorption and entry was obtained from Escherichia coli using recombinant protein expression and purification. SDS-PAGE revealed an approximate size of 30 kDa for N1N2. Dot blot and transmission electron microscopy analyses revealed that the protein bound to the host V. cholerae but not to non-host E. coli K-12 cells. Next, we used amine-coupling to develop a novel recombinant N1N2 (rN1N2)-functionalized SPR biosensor by immobilizing rN1N2 proteins on gold substrates and using SPR to monitor the binding kinetics of the proteins with target bacteria. We observed rapid detection of V. cholerae in the range of approximately 10³ to 10⁹ CFU/ml but not of E. coli at any tested concentration, thereby confirming that the biosensor exhibited differential recognition and binding. The results indicate that the novel biosensor can rapidly monitor a target pathogenic microorganism in the environment and is very useful for monitoring food safety and facilitating early disease prevention.

• 센서학회지
• /
• 제22권2호
• /
• pp.144-149
• /
• 2013

In this research, we developed a biosensor to detect lung cancer-specific biomarker using Anodic Aluminum Oxide (AAO) chip based on interference and nano surface plasmon resonance (nanoSPR). The nano-porous AAO chip was fabricated $2{\mu}m$ of pore-depth by two-step anodizing method for surface uniformity. NanoSPR has sensitivity to the refractive index (RI) of the surrounding medium and also provides simple and label-free detection when specific antibodies are immobilized to the Au-deposited surface of nano-porous AAO chip. To detect the lung cancer-specific biomarker, antibodies were immobilized on the surface of the chip by Self Assembled Monolayer (SAM) method. Since then lung cancer-specific biomarker was applied atop the antibodies immobilized layer. The specific reaction of the antigen-antibody contributed to the change in the refractive index that cause shift of resonance spectrum in the interference pattern. The Limit of Detection (LOD) was 1 fg/ml by using our nano-porous AAO biosensor chip.

• Food Science and Biotechnology
• /
• 제17권3호
• /
• pp.547-552
• /
• 2008

The detection of carbamate (carbofuran, carbaryl, benfracarb, thiodicarb, and methomil) and organophosphate (diazinon, cadusafos, ethoprofos, parathion-methyl, and chlorpyrifos) pesticide residues with very low detection limits was carried out using surface plasmon resonance (SPR) based equipment. The capacity to develop a portable SPR biosensor for food safety was also investigated. The applied ligand for the immunoassays was polyclonal goat anti-rabbit immunoglobulin (IgG) peroxidase conjugate. Concentration tests using direct binding assays showed the possibility of quantitative analysis. For ligand fishing to find a proper antibody to respond to each pesticide, acetylcholinesterase (AChE), and glutathione-S-transferase (GST) were tested. The reproducibility and precision of SPR measurements were evaluated. With this approach, the limit of detection for pesticide residues was 1 ng/mL and analysis took less than 11 min. Thus, it was demonstrated that detecting multi-class pesticide residues using SPR and IgG antibodies provides enough sensitivity and speed for use in portable SPR biosensors.

• 한국광학회지
• /
• 제24권2호
• /
• pp.86-91
• /
• 2013

본 논문에서는 근접장-분자반응 간의 중첩을 이용한 표면 플라스몬 공명 (SPR) 바이오센서의 측정감도 평가방법을 연구하였다. 전달행렬 방법을 사용하여 다양한 형태의 중첩적분으로 정의된 광학자취 값을 계산하였고, 샌드위치 및 역샌드위치 면역글로뷸린 (IgG) 어세이에 대해서 실험적으로 측정된 수치와 비교하였다. 이론 및 실험적인 결과와의 비교를 통하여 접선 방향의 전기장을 사용한 광학자취의 경우 그 연속성으로 말미암아 가장 높은 상관계수를 얻을 수 있었으며 이때 광학자취와 측정감도 사이에 97% 이상의 높은 상관계수가 존재함을 보았다. 이러한 상관관계는 SPR 바이오센서의 측정 감도에 관한 메커니즘을 분명하게 설명하며, 분자 스케일 감도를 가지는 SPR 바이오센서 개발에 기여하게 될 것이다.