• 한국표면공학회지
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• 제37권4호
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• pp.215-219
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• 2004

We fabricated protein chip plates coated with silane carboxylate. The silane compound was immobilized by hydrogen bond and/or other chemical bonds on the surface of the plate. The plates were then prepared by binding $Ni^{2+}$ to surfaces terminated with silane carboxylate groups. The carboxylic acid surface was generated by chemical oxidation of the terminal double-bond functions of the silane-deposited layer. The $Ni^{2+}$ ions on the surface reacted readily to His-tagged proteins. A significant increase in His-tagged protein adsorption was achieved on the surface terminated with silane carboxylate with longer alkyl chain, suggesting better availability of these protein chip plates for proteomic studies.

• 한국표면공학회지
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• 제37권2호
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• pp.76-79
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• 2004

Nickel chloride coated protein chip plate was developed by using a spin coating method. The ability of histidine tagged protein adsorption was investigated at various solvents. The surface of plate has a large aggregated nickel complex with high density in water. However, the surface of plate has a very small size of aggregated nickel complex with low density in isopropanol. The ability of protein adsorption decreased as increasing the size of alkyl chain in various alcohol solvents. The mechanism on the ability of protein adsorption at the plate surface is discussed.

• Journal of Microbiology and Biotechnology
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• 제31권8호
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• pp.1183-1189
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• 2021

Autodisplay of a multimeric protein complex on a cell surface is limited by intrinsic factors such as the types and orientations of anchor modules. Moreover, improper folding of proteins to be displayed often hinders functional cell surface display. While overcoming these drawbacks, we ultimately extended the applicability of the autodisplay platform to the display of a protein complex. We designed and constructed a cell surface attachment (CSA) system that uses a non-covalent protein-protein interaction. We employed the high-affinity interaction mediated by an orthogonal cohesin-dockerin (Coh-Doc) pair from Archaeoglobus fulgidus to build the CSA system. Then, we validated the orthogonal Coh-Doc binding by attaching a monomeric red fluorescent protein to the cell surface. In addition, we evaluated the functional anchoring of proteins fused with the Doc module to the autodisplayed Coh module on the surface of Escherichia coli. The designed CSA system was applied to create a functional attachment of dimeric α-neoagarobiose hydrolase to the surface of E. coli cells.

• 한국표면공학회지
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• 제40권2호
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• pp.98-102
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• 2007

[ $NiCl_2$ ] and poly-L-lysine coated protein chip plates have been fabricated using a spin coating system. Water has been used as solvent and scratching effects on glass slides and ITO have been investigated. We also observed the surface properties of $NiCl_2$ and poly-L-lysine coated slides by using PSA(Particle size analyzer) and AFM(Atomic force microscope). The AFM results imply that the surface patterns created in the spin coating system determine the protein adsorption. Adsorption of histidine-tagged KRS proteins immobilized on glass slides and ITO was analyzed using a BAS image system. The results suggest that the scratching effect was increased ability of protein adsorption.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.47-47
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• 2005

There are many different surface technologies currently applied for preparation of protein chips. However, it requires innovative surface chemistry for capture proteins to be immobilized on chip surface keeping their conformation and activity intact and their orientation right, while they bind tightly and densely in a given array spot. Proteogen has developed 'ProteoChip BP' coated with novel proprietary linker molecules $(ProLinker^{TM})$ for efficient and robust immobilizations of capture proteins by improving surface properties of molecular captures. It was demonstrated that $ProLinker^{TM}$ gave the best surface performance in preparation of protein microarray chip base plates among others currently available on the market. In particular, the $ProLinker^{TM}-based$ surface chemistry has demonstrated to provide excellent performance in preparation of 'Antibody Chip' for analysis of biomarkers as well as proteome expression profiles. The linker molecule has also shown to be well applicable for development of biosensors and micro-beads as well as protein microarray and nano-array. ProteoChip BP can be used either for preparation of high-density array by using a microarrayer or for preparation of 'Well-on-a-Chip' with low density array, which is better applicable for quantitative analysis of biomarkers or protein-protein interactions. The biomarker assay can be performed either by direct or sandwich methods of fluorescence immunoassay. Application of ProteoChip BP has been well demonstrated by the extensive studies of 1) tumor-marker assays, 2) new drug screening by using 'Integrin Chip' and 3) protein expression profile analysis. Some of experimental results will be presented.

• Membrane and Water Treatment
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• 제1권2호
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• pp.103-120
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• 2010

Surface modification of microfiltration and ultrafiltration membranes has been widely used to improve the protein adsorption resistance and permeation properties of hydrophobic membranes. Several surface modification methods for converting conventional membranes into low-protein-binding membranes are reviewed. They are categorized as either physical modification or chemical modification of the membrane surface. Physical modification of the membrane surface can be achieved by coating it with hydrophilic polymers, hydrophilic-hydrophobic copolymers, surfactants or proteins. Another method of physical modification is plasma treatment with gases. A hydrophilic membrane surface can be also generated during phase-inverted micro-separation during membrane formation, by blending hydrophilic or hydrophilic-hydrophobic polymers with a hydrophobic base membrane polymer. The most widely used method of chemical modification is surface grafting of a hydrophilic polymer by UV polymerization because it is the easiest method; the membranes are dipped into monomers with and without photo-initiators, then irradiated with UV. Plasma-induced polymerization of hydrophilic monomers on the surface is another popular method, and surface chemical reactions have also been developed by several researchers. Several important examples of physical and chemical modifications of membrane surfaces for low-protein-binding are summarized in this article.

• Applied Science and Convergence Technology
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• 제24권1호
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• pp.1-8
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• 2015

Protein immobilization on a gold surface plays an important role in the usefulness of biosensors that utilize gold-coated surfaces such as surface plasmon resonance (SPR), quartz crystal microbalance (QCM), etc. For developing high performance biosensors, it is necessarily required that immobilized proteins must remain biologically active. Loss of protein activity and maintenance of its stability on transducer surfaces is directly associated with the choice of immobilization methods, affecting protein-protein interactions. During the past decade, a variety of strategies have been extensively developed for the effective immobilization of proteins in terms of the orientation, density, and stability of immobilized proteins on analytical devices operating on different principles. In this review, recent advances and novel strategies in protein immobilization technologies developed for biosensors are briefly discussed, thereby providing an useful information for the selection of appropriate immobilization approach.

• 공업화학
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• 제30권3호
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• pp.379-383
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• 2019

The capability of detecting amyloid beta 42 ($A{\beta}42$), a biomarker of Alzheimer's disease, using a thiolated protein A-functionalized bimetallic surface plasmon resonance (SPR) chip was investigated. An optimized configuration of a bimetallic chip containing gold and silver was obtained through calculations in the intensity measurement mode. The surface of the SPR bimetallic chip was functionalized with thiolated protein A for the immobilization of $A{\beta}42$ antibody. The response of the thiolated protein A-functionalized bimetallic chip to $A{\beta}42$ in the concentration range of 50 to 1,000 pg/mL was linear. Compared to protein A without thiolation, the thiolated protein A resulted in greater sensitivity. Therefore, the thiolated protein A-functionalized bimetallic SPR chip can be used to detect very low concentrations of the biomarker for Alzheimer's disease.

• 센서학회지
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• 제20권6호
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• pp.434-438
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• 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.

• 한국진공학회지
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• 제17권6호
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• pp.549-554
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• 2008

수소 플라즈마 처리된 유리 기판에 스핀 코팅 시스템을 이용하여 nickel chloride를 코팅하여 단백질칩 플레이트를 제조하였다. 다양한 플라즈마 처리 시간대에서 histidine tagged 단백질의 부착 능력 특성을 연구하였다. 유리 기판 표면에서 nickel chloride와 단백질 특성을 particle size analysis를 이용하여 관찰하였고, 단백질의 부착 능력 정도를 bio imaging analyzer system으로 측정하였다. 실험 결과에 따르면, 플라즈마 처리 시간이 증가할수록 단백질 부착 능력은 감소하는 것으로 나타났다. 기판 표면에서의 단백질 부착능력 특성에 관한 mechanism은 본문의 결과 및 토의에서 논의되었다. 플라즈마 처리된 단백질칩 기판에 대한 표면 안정화는 바이오센서 시장에서 큰 관심을 끌 것으로 기대된다.