• Journal of Microbiology and Biotechnology
• /
• 제16권8호
• /
• pp.1229-1235
• /
• 2006

A lab-on-a-chip (LoC) was designed for simultaneous monitoring of microorganisms, antibiotic residues, somatic cells, and pH in raw milk. The LoC was fabricated from polydimethylsiloxane (PDMS) using microelectromechanical system (MEMS) technology, which consisted of two parts; a protein array and microchannel. The protein array was fabricated by immobilizing five types of antibodies corresponding to two microorganisms, two antibiotic residues, and somatic cells. A sol-gel film was deposited on a glass substrate to immobilize the antibodies. The target analytes in raw milk could be bound with the corresponding antibody by an immunoreaction, and the antigen-antibody complex was detected using fluorescence microscopy. SNARF-dextran was used as a pH indicator, and the SNARF-entrapped hydrogel was attached to the microchannel in the chip. After injecting the milk sample into the channel, the pH was measured by monitoring the change in fluorescence intensity by fluorescence microscopy. The on-chip simultaneous assay of two microorganisms (E. coli O157:H7 and Streptococcus agalactiae), two antibiotic residues (penicillin G and dihydrostreptomycin), and neutrophils was successfully accomplished using the proposed LoC system.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권9호
• /
• pp.3709-3713
• /
• 2015

Background: Many factors, including molecular ones, were demonstrated to be associated with long-term prognosis of hepatocellular carcinoma (HCC). Thus far, the expression and clinicopathologic and prognostic significance of the carboxyl terminus of Hsp70-interacting protein (CHIP) in B-type hepatitis virus (HBV)-related HCC remain unknown. Materials and Methods: CHIP expression was detected by immunohistochemical staining of surgical samples from 79 patients with HCC with HBsAg positivity. In addition, correlations with clinicopathologic parameters and patient survival were evaluated. Results: It was found that positive CHIP staining was observed in tumor, but not non-tumor, tissues. High expression of CHIP was significantly related to larger tumor size, with marginally significant associations noted for presence of portal vein invasion and higher serum a-fetoprotein level. In addition, univariate analysis showed that high CHIP expression was a powerful predictor for dismal overall and disease-free survival. However, independent prognostic implications of CHIP were not proven in multivariate Cox regression test. Conclusions: CHIP is overexpressed in HBV-related HCC and is associated with unfavorable biological behavior as well as poor prognosis. However, its prognostic role needs to be further validated.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제11권2호
• /
• pp.146-153
• /
• 2006

A passive microfluidic delivery system using hydrophobic valving and pneumatic control was devised for microfluidic handling on a chip. The microfluidic metering, cutting, transport, and merging of two liquids on the chip were correctly performed. The error range of the accuracy of microfluid metering was below 4% on a 20 nL scale, which showed that microfluid was easily manipulated with the desired volume on a chip. For a study of the feasibility of biochemical reactions on the chip, a single enzymatic reaction, such as ${\beta}-galactosidase$ reaction, was performed. The detection limit of the substrate, i.e. fluorescein $di-{\beta}-galactopyranoside$ (FDG) of the ${\beta}-galactosidase$ (6.7 fM), was about 76 pM. Additionally, multiple biochemical reactions such as in vitro protein synthesis of enhanced green fluorescence protein (EGFP) were successfully demonstrated at the nanoliter scale, which suggests that our microfluidic chip can be applied not only to miniaturization of various biochemical reactions, but also to development of the microfluidic biochemical reaction system requiring a precise nano-scale control.

• 한국어병학회지
• /
• 제27권2호
• /
• pp.99-105
• /
• 2014

Biosensors consist of biochemical recognition agents like antibodies immobilized on the surfaces of transducers that change the recognition into a measurable electronic signal. Here we report a piezoelectric immunosensor made to detect Vibrio vulnificus. A 9MHz AT-cut piezoelectric wafer attached with two gold electrodes of 5mm diameter was used as the transducer of the QCM biosensor with a reproducibility of ${\pm}0.1Hz$ in frequency response. We have tried different approaches to immobilize antibody on the sensor chip. Concerning the orientation of antibody for the best antigen binding capacity, the antibody was immobilized by specific binding to protein G or by cross-linking through hydrazine. In addition, protein G was cross-linked on glutaraldehyde activated immine layer (PEI) or EDC/NHS activated sulfide monolayer (MPA). PEI was found to be more effective to immobilize protein G following glutaraldehyde activation than MPA. However, hydrazine chip showed a better capability to immobilize more IgG than protein G chip and a higher sensitivity. The sensor system was able to detect V. vulnificus in dose dependent manner and was able to detect bacterial cells within 5 minutes by monitoring frequency shifts in real time. The detection limit can be improved by preincubation to enrich the bacterial cell number.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제11권5호
• /
• pp.455-461
• /
• 2006

In this study, we have described a method for the fabrication of a protein chip on silicon substrate using hydrophobic thin film and microfluidic channels, for the simultaneous detection of multiple targets in samples. The use of hydrophobic thin film provides for a physical, chemical, and biological barrier for protein patterning. The microfluidic channels create four protein patterned strips on the silicon surfaces with a high signal-to-noise ratio. The feasibility of the protein chips was determined in order to discriminate between each protein interaction in a mixture sample that included biotin, ovalbumin, hepatitis B antigen, and hepatitis C antigen. In the fabrication of the multiplexed assay system, the utilization of the hydrophobic thin film and the microfluidic networks constitutes a more convenient method for the development of biosensors or biochips. This technique may be applicable to the simultaneous evaluation of multiple protein-protein interactions.

• BioChip Journal
• /
• 제12권4호
• /
• pp.317-325
• /
• 2018

This paper investigated the effects of ionic strength in the medium on a preconcentrator for a protein sample with low concentration. The preconcentration chip was designed and fabricated using a polydimethylsiloxane replica through standard lithophotography. A glass substrate is silanized prior to functionalizing the nanoparticles for self-assembly at a designed region. Due to the overlap of electrical double layers in a nanofluidic channel, a concentration polarization effect can be achieved using an electric field. A nonlinear electrokinetic flow is induced, resulting in the fast accumulation of proteins in front of the induced ionic depletion zone, so called exclusion-enrichment effect. Thus, the protein sample can be driven by electroosmotic flow and accumulated at a specific location. The chip is used to collect fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) diluted in phosphate-buffered saline (PBS) buffer solution. Different concentrations of the buffer media were studied herein. Fluorescence intensity images show that the buffer concentration of 4 mM is more appropriate than all the other ones. The sample of FITC-BSA with an initial concentration of $10{\mu}M$ in the 4 mM PBS solution increases its concentration at the desired region by up to 50 times within 30 min, demonstrating the results in this investigation.

• 한국진공학회:학술대회논문집
• /
• 한국진공학회 2006년도 제31회 학술논문발표회 초록집
• /
• pp.42-42
• /
• 2006

• 센서학회지
• /
• 제20권6호
• /
• pp.434-438
• /
• 2011

A portable surface plasmon resonance(SPR) based immunosensor using thiolated protein G and protein G was developed for the detection of immunoglobulin G(IgG). The protein G has specific affinity with Fc fragment of IgG and was thiolated by 2-Iminothiolane for introduction of thiol groups. Anti-IgG, bovine serum albumin(BSA), and IgG have been sequently injected after surface modification of gold sensor chip with protein G and thiolated protein G. The output signal was increased with the injection of each protein and the actual signal was measured by subtracting signal of reference channel from signal of sample injected channel. The experimental results showed the higher detection capability of IgG using thiolated protein G compared with protein G. From these results, we can conclude that the current surface modification technique and the portable SPR sensor system can be applied to various immunosensors for diagnosis.

• Korean Chemical Engineering Research
• /
• 제44권1호
• /
• pp.1-9
• /
• 2006

생물체를 구성하는 세포의 기능과 구성요소 간 상호작용 메커니즘을 인공적으로 모방하여 바이오물질 박막으로 구성된 바이오소자는 의료 진단, 신약 스크리닝, 전자소자, 생물공정, 환경오염 물질 측정 등 다양한 산업 분야에 응용되고 있다. 단백질, DNA, 바이오색소, 세포 등의 생체물질을 칩 상에 고집적으로 배열하여 구성된 바이오 소자로서 바이오 전자소자(생물분자 광다이오드, 바이오 정보저장소자, 바이오 전기발광 소자), DNA칩, 단백질칩, 및 세포칩 등이 개발되어 오고 있다. 생체물질 고정화 기술, 마이크로 및 나노수준의 패터닝기술, 소자 구성 기술, 바이오 멤스 기술의 융합을 통해 바이오소자는 구현되며, 최근에는 나노기술의 적용에 의하여 나노바이오소자도 구현이 가능하다. 본 논문에서는 현재까지 개발된 다양한 바이오소자의 제작 기술과 응용에 대하여 소개하고 향후의 발전 방향에 대하여 다룬다.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2001년도 추계학술발표대회
• /
• pp.859-862
• /
• 2001

For the development of preferable immunosensor and protein chip, the viologen Langmuir-Blodgett (LB) multilayer was fabricated on the surface, and then protein A was adsorbed on the proposed viologen LB film by electrostatic attractive force. The Immunoglobulin G (IgG) labeled with fluorescence marker was self-assembled on the fabricated protein A film. The topographies of the deposited films were investigated by using atomic force microscope (AFM). The immobilization of IgG was verified by fluorescence spectrum. Such structures can be used as sublayers for various kinds of IgG immobilization toward immunosensors and protein chip.