• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.859-862
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• 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.

• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.65-72
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• 2006

Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.1
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• pp.19-23
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• 1998

We enhanced the sensitivity of surface plasmon resonance biosensor by the conversion of the real-time direct binding immunoassay into the sandwich immunoassay, in which colloidal gold particles coated with anti-mouse IgG was used. By the immobilization of anti-mouse IgG onto the carboxymethyl dextran surface of thin gold film, the direct binding of analyte(mouse IgG) onto the sensor chip, and the injection of colloidal gold particles coated with anti-mouse IgG, about 100 times of sensitivity enhancement was obtained. This result suggests that nanoparticles, which has a high refractive index, homogeneous ultrafine structure and capability of size control, would be applicable for the detection of very small quantity of biomaterial.

• 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.

• Journal of Sensor Science and Technology
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• v.20 no.4
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• pp.226-229
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• 2011

Label-free detection of biomolecular interactions was performed using BioFET(Biologically sensitive Field-Effect Transistor) and SPR(Surface Plasmon Resonance). Qualitative information on the immobilization of an anti-IgG and antibody-antigen interaction was gained using the SPR analysis system. The BioFET was used to explore the pI value of the protein and to monitor biomolecular interactions which caused an effective charge change at the gate surface resulting in a drain current change. The results show that the BioFET can be a useful monitoring tool for biomolecular interactions and is complimentary to the SPR system.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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• pp.71-80
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• 1990

New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.

• Analytical Science and Technology
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• v.30 no.1
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• pp.49-56
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• 2017

Here, we demonstrate surface functionalization of Au chips with 4-(carboxymethyl)aniline (CMA) and amine-terminated polyamidoamine (PAMAM) dendrimers for immobilization of antibodies on the Au surfaces. Use of the functionalization strategy led to high surface density of the immobilized antibodies on the Au chips. Specifically, we found that the functionalization of Au chips with CMA and amine-terminated $6^{th}$ generation PAMAM dendrimers allowed immobilization of immunoglobulin (IgG) antibodies with high surface density, which is 5 times higher than that obtained with Au surfaces functionalized with CMA and ethylenediamine.

• Korean Chemical Engineering Research
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• v.49 no.2
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• pp.224-229
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• 2011

In this study, we have developed shear horizontal(SH) surface acoustic wave(SAW) sensors for the detection of immunoglobulin G(IgG) on the gold coated delay line of SH-SAW devices. As the result of the experiment, we could uniformly immobilize anti-MIgG(mouse IgG) conjugate on the surface of gold. When displaying results of immobilization on the surface of gold using G-anti MIgG conjugate and blocking buffer in frequency shift, G-anti MIgG conjugate showed frequency shift of 75.1 kHz in the initial frequency, and blocking buffer showed frequency shift of 215.7 kHz. When various concentrations of MIgG was added in 100MHz type sensor, the sensor showed 46.3, 127.45, 161.21 and 262.39 kHz frequency shift at 25, 50, 75 and 100 ${\mu}g$ MIgG concentration, respectively.

• Bulletin of the Korean Chemical Society
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• v.32 no.6
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• pp.1975-1979
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• 2011

The modification of a gas diffusion layer (GDL), a vital component in polymer electrolyte fuel cells, is described here for use in the electrochemical detection of antibody-antigen biosensors. Compared to other substrates (gold foil and graphite), mouse anti-rHBsAg monoclonal antibody immobilized on gold-coated GDL (G-GDL) detected analytes of goat anti-mouse IgG antibody-ALP using a relatively low potential (-0.0021 V vs. Ag/AgCl 3 M NaCl), indicating that undesired by-reactions during electrochemical sensing should be avoided with G-GDL. The dependency of the signal against the concentration of analytes was observed, demonstrating the possibility of quantitative electrochemical biosensors based on G-GDL substrates. When a sandwich method was employed, target antigens of rHBsAg with a concentration as low as 500 ng/mL were clearly measured. The detection limit of rHBsAg was significantly improved to 10 ng/mL when higher concentrations of the 4-aminophenylphosphate monosodium salt (APP) acting on substrates were used for generating a redox-active product. Additionally, it was shown that a BSA blocking layer was essential in improving the detection limit in the G-GDL biosensor.