• Asian Pacific Journal of Cancer Prevention
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• v.16 no.12
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• pp.5115-5118
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• 2015

Background: The $p16^{INK4a}$ is a protein that expressed in Liquid-based cervical cytology specimens and has been proved link to cervical cancer. The $p16^{INK4a}$ could be detection by piezoelectric immunosensor and the immobilization of the $p16^{INK4a}$ antibody influence the sensitivity of the piezoelectric immunosensor. Materials and Methods: $5{\mu}L$ mouse polyclonal antibody against $p16^{INK4a}$ was bound onto the surface of immonosensor through two methods. (directly immobilized method; protein A method). Absorb of the $p16^{INK4a}$ antibody on the surface of immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changes recorded showed a better reproducibility. The activity of the immobilization antibody with the directly method and protein A method was tested with $p16^{INK4a}$ antigen. Results: The resonant frequency for different antibody immobilization methods were different, and the sensitivity for $p16^{INK4a}$ detection also different. Conclusions: The protein A method was found to be much more better than the directly method for the immobilization of the p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein A method created more reproducible and stable immobilization antibody layers with p16INK4A antigen.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.701-705
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• 2003

Performance of an immunosensor can usually be assessed in terms of its analytical sensitivity and specificity. Sensitivity, i.e., the detection limit of analyte, is particularly determined by the amount of analyte molecules bound to the capture antibody immobilized onto a solid surface. In order to increase the binding complexes, we have investigated an immobilization method of antibody allowing for a molecular arrangement of the protein on a selective surface of a nano-patterned solid substrate. This has not been accomplished only by a surface treatment with a chemical, but also by fragmentation of immunoglobulin. Such approach would offer a protocol of antibody immobilization for the construction of nano-immunosensor and eventually improve the sensitivity of detection.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.112-117
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• 2004

The performance of an immuno-analytical system can be assessed in terms of its analytical sensitivity, i.e., the detection limit of an analyte, which is determined by the amount of analyte molecules bound to the capture antibody that has been immobilized onto a solid surface. To increase the number of the binding complexes, we have investigated a site-directed immobilization of an antibody that has the ability to resolve a current problem associated with a random arrangement of the insolubilized immunoglobulin. The binding molecules were chemically reduced to produce thiol groups that were limited at the hinge region, and then, the reduced products were coupled to biotin. This biotinylated antibody was bound to a streptavidin-coated surface via the streptavidin-biotin reaction. This method can control the orientation of the antibody molecules present on a solid surface and also can significantly reduce the possibility of steric hindrance in the antigen-antibody reactions. In a two-site immunoassay, the introduction of the site-directly immobilized antibody as the capture enhanced the sensitivity of analyte detection approximately 10 times compared to that of the antibody randomly coupled to biotin. Such a novel approach would offer a protocol of antibody immobilization in order for the possibility of constructing a high performance immunochip.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.4
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• pp.241-244
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• 2004

Protein A molecular thin film was fabricated as a platform of antibody-based biosensor. For the immobilization of the protein A thin film, a viologen multilayer was built up using the Langmuir-Blodgett (LB) technique, and then, protein A was adsorbed on the viologen LB film by an electrostatic interaction force, which was formed as a hetero-film structure. For the deposition of viologen, surface pressure area ($\pi$-A) isotherm was investigated. The fabricated protein A-viologen hetero LB film was investigated using atomic force microscopy (AFM). Using the developed molecular film, antibody immobilization and fluorescence measurement was carried out.

• Journal of Biosystems Engineering
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• v.34 no.4
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• pp.254-259
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• 2009

Conventional methods for pathogen detection and identification are labor-intensive and take several days to complete. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella typhimurium. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on either avidin-biotin binding or self assembled monolayer (SAM) on the surface of the IME to form an active sensing layer. To evaluate effect of antibody immobilization methods on sensitivity of the sensor, detection limit of the biosensor was analyzed with Salmonella samples innoculated in phosphate buffered saline (PBS) or food extract. The impedimetric biosensor based on SAM immobilization method produced better detection limit. The biosensor could detect 107 CFU/mL of Salmonella in pork meat extract. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.

• Bulletin of the Korean Chemical Society
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• v.24 no.1
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• pp.70-74
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• 2003

The heterogeneous bioluminescence immunoassay for digoxin was developed using photoprotein, native aequorin as a label and the site-directed immobilization technique based on avidin/biotin interaction. Aequorin is a bioluminescence protein, originally isolated from the jellyfish Aequoria Victoria and an attractive label in analytical applications because of sensitive detection due to virtually no background bioluminescent signal. Digoxin is a cardioactive drug, and its therapeutic level in serum is at low concentration with very narrow therapeutic index. The aequorin-digoxigenin conjugates were synthesized by the N-hydroxysuccinimide ester method and characterized in terms of bioluminescent residual activity. The resulting dose-response curve shows that the detection limit is $1.0\;{\times}\;10^{-10}\;M$ and a dynamic range is three orders of magnitude, which was obtained by $1.0\;{times}\;10^{-10}\;M$ conjugate and 0.9 μg/mL anti-digoxin antibody. Three structurally similar molecules to digoxin were examined for their cross-reactivity. None of these three compounds showed any crossreactivity with digoxin antibody employed in this study. Standard amounts of digoxin corresponding to the therapeutic range were spiked into the each serum solution. Study of the serum matrix effect indicated that correlation coefficient shows good agreement between luminescence light intensity between in buffer and in serum.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1169-1173
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• 2006

Imaging ellipsometry (IE) for detection of binding of Escherichia coli O157:H7 (E. coli O157:H7) to an immunosensor is reported. A protein G layer, chemically bound to a self-assembled layer of 11-mercaptoundecanoic acid (11-MUA), was adopted for immobilization of monoclonal antibody against E. coli O157:H7 (Mab). The immobilization of antibody was investigated using surface plasmon resonance. To fabricate antibody spots on a gold surface, protein G solution was spotted onto the gold surface modified with an 11-MUA layer, followed by immobilizing Mab on the protein G spot. Ellipsometric images of the protein G spot, the Mab spot, and Mab spots with binding of E. coli O157:H7 in various concentrations were acquired using the IE system. The change of mean optical intensity of the Mab spots in the ellipsometric images indicated that the lowest detection limit was $10^3$CFU/ml for E. coli O157:H7. Thus, IE can be applied to an immunosensor for detection of E. coli O157:H7 as a detection method with the advantages of allowing label-free detection, high sensitivity, and operational simplicity.

• Bulletin of the Korean Chemical Society
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• v.27 no.4
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• pp.479-483
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• 2006

We developed a microfluidic immunoassay platform for the detection of various analytes such as bacterial pathogen by packing antibody-immobilized glass beads in spatially-isolated microchambers on a microfluidic device. Primary amines of antibody were covalently conjugated to carboxyl-terminated glass beads previously treated with aminosilane followed by glutaraldehyde. Through this covalent binding, up to 905 $\mu$g immunoglobulin G (IgG) per gram of glass beads was immobilized. For application, glass beads attaching antibody specific to Escherichia coli O157:H7, a foodborne pathogen, were packed into a microfluidic device and used for the detection of the serotype. This prototype immunoassay device can be used for the simultaneous detection of multiple analytes by sequentially packing different-sized glass beads attaching different antibody in discrete microchambers on a single microfluidic device.

• Korean Journal of Food Science and Technology
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• v.31 no.1
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• pp.1-6
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• 1999

A piezoelectric (PZ) biosensor system detecting Salmonella spp. was developed. The system consisted of an oscillator, a frequency counter and an antibody-immobilized quartz crystal. An anti-Salmonella antibody was immobilized on one gold. surface of the quartz crystal with protein A. Salmonella detection was made by measuring resonant frequency shift owing to a mass change by specific binding of microbial cells to the gold surface of the PZ crystal. The PZ antibody sensor was operated optimally at 0.1M phosphate buffer, pH 7.2 and $35^{\circ}C$. The sensor was quite specific to Salmonella spp. The obtained frequency shift was correlated with the Salmonella concentration in the range of $10^5{\sim}10^6\;CFU/mL$. The frequency shift increased further by addition of polystyrene beads. The Salmonella detection which is indicated by a steady-state microbial adsorption to the quartz crystal was accomplished within 50min.

• Polymer(Korea)
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• v.33 no.6
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• pp.602-607
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• 2009

A polyurethane (PU) surface enabling in vivo endothelialization via endothelial progenitor cell (EPC) capture was prepared for cardiovascular applications. To introduce CD34 monoclonal antibody (mAb) inducing EPC adhesion onto a surface, poly (poly (ethylene glycol) acrylate-co-butyl methacrylate) and poly (PEGA-co-BMA) were synthesized and then coated on a surface of PU, followed by immobilizing CD34 mAb. $^1H$-NMR analysis demonstrated that poly(PEGA-co-BMA) copolymers with a desired composition were synthesized. Poly(PEGA-co-BMA)-coated PU was much more effective for the immobilization of CD34 mAb, comparing with PEG-grafted PU prepared in our previous study, as demonstrated by that surface density and activity of CD34 mAb increased over 32 times. Physico-chemical properties of modified PU surfaces were characterized by X-ray photoelectron spectroscopy (XPS), water contact angle, and atomic force microscopy (AFM). The results demonstrated that the poly(PEGA-co-BMA) coating was effective for CD34 mAb immobilization and feasible for applying to cardiovascular biomaterials.