• Title, Summary, Keyword: avidin-biotin

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A Study on Surface Modification of Nanorod Electrodes for Highly Sensitive Nano-biosensor (고감도 나노-바이오센서를 위한 나노로드 전극 표면 개질에 관한 연구)

  • Lee, Seung Jun
    • Applied Chemistry for Engineering
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    • v.27 no.2
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    • pp.185-189
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    • 2016
  • Among many kinds of bioaffinity sensors, the avidin-biotin system has been widely used in a variety of biological applications due to the specific and high affinity interaction of the system. In this work, gold nanorods with high surface area were explored as electrodes in order to amplify the signal response from the avidin-biotin interaction which can be further utilized for avidin-biotin biosensors. Electrochemical performance of electrodes modified with nanorods and functionalized with avidin in response to interactions with biotin at various concentrations using $[Fe(CN)_6]^{3-/4-}$ couple as the redox probe were investigated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). A very low biotin concentration of less than 1 ng/mL could be detected using the electrodes modified with nanorods.

Characterization of biotin-avidin recognition system constructed on the solid substrate

  • Lim, Jung-Hyurk
    • Analytical Science and Technology
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    • v.18 no.6
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    • pp.460-468
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    • 2005
  • The biotin-avidin complex, as a model recognition system, has been constructed through N-hydroxysuccinimide(NHS) reaction on a variety of substrates such as a smooth Au film, electrochemically roughened Au electrode and chemically modified mica. Stepwise self-assembled monolayers (SAMs) of biotin-avidin system were characterized by surface-enhanced resonance Raman scattering (SERRS) spectroscopy, atomic force microscopy (AFM) and surface plasmon resonance (SPR). A strong SERRS signal of rhodamine tags labeled in avidin from the SAMs on a roughened gold electrode indicated the successful complex formation of stepwise biotin-avidin recognition system. AFM images showed the circular shaped avidin aggregates (hexamer) with ca. $60{\AA}$ thick on the substrate, corresponding to one layer of avidin. The surface coverage and concentration of avidin molecules were estimated to be 90% and $7.5{\times}10^{-12}mol/cm^2$, respectively. SPR technique allowed one to monitor the surface reaction of the specific recognition with high sensitivity and precision.

Effect of Avidin and Biotin in Attachment of Human Adipose Stem Cells to Micronized Acellular Dermal Matrix (지방줄기세포가 무세포 분쇄진피기질(Acellular micronized dermal matrix) 부착에 있어 Avidin과 Biotin의 효과)

  • You, Gyeol;Rhie, Jong Won;Lim, Jin Soo
    • Archives of Plastic Surgery
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    • v.35 no.1
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    • pp.1-6
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    • 2008
  • Purpose: In tissue engineering, it is important that the scaffolds have high affinity with cells for making efficient use of cells. The authors studied the binding affinity of human adipose stem cells(ASCs) to micronized acellular dermal matrix(alloderm) using biotin and avidin linkages.Methods: Human ASCs were harvested from adipose tissue obtained by abdominoplasty. ASCs($1{\times}10^4$, $5{\times}10^4$, $1{\times}10^5$, $5{\times}10^5$, $1{\times}10^6$, $5{\times}10^6$ cells) were attached to micronized alloderm(1mg) in three groups; 1) control group in which no ASCs and alloderm was treated; 2) serum group in which alloderm was exposed to fetal bovine serum; and 3) biotin group in which biotinylated cells were attached to biotinylated alloderm. The binding affinities were determined 1 day after making ASC-alloderm complexes. The proliferation rates were determined by XTT assays in 4, 7, 14, and 21 days and scanning electron microscopic examination was performed in 7 and 21 days after culture of ASC-alloderm complexes.Results: The binding affinities of the biotin group were significantly increased in all cell concentrations. Maximum binding affinity was observed at $5{\times}10^4/mg$ of micronized dermal matrix in biotin group. The viabilities were lowest in biotin group in contrast to binding affinity, but the difference was not significant. SEM showed well attachment of cells to micronized dermal matrix in all groups. Conclusion: The use of avidin/biotin facilitated human ASCs attaching to micronized acellular dermal matrix. This attachment would not disturb adipose stem cells viabilities. The present study suggests that avidin/ biotin can be used as making efficient use of cells in adipose tissue engineering.

Enzyme Sensors Modified with Avidin/Biotin Systembased Protein Multilayers

  • Anzai, Jun-Ichi;Du, Xiao-Yan;Hoshi, Tomonori;Suzuki, Yasuhiro;Takeshita, Hiroki;Osa, Tetsuo
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.591-596
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    • 1995
  • Enzyme multilayers composed of avidin and biotin-labeled enzymes were prepared on the surface of electrode, through a strong affinity between avidin and biotin (binding constant: ca $10^{15} M^{-1}$). The enzyme multilayers were useful for the improvement of the performance characteristies of enzyme sensors. The output current of the enzyme sensors depended linearly on the number of enzyme layers deposited. Thus, lactate oxidase (LOx) and alcohol oxidase (AlOx) were deposited after being modified with biotin for constructing enzyme sensors sensitive to L-lactate and ethanol respectively. It was also possible to deposit two different kinds of enzymes successively in a single multilayer. The glucose oxidase (GOx) and ascorbate oxidase (AsOx) were built into a multilayer structure on a Platinum electrode. The GOx, AsOx multilayer-modified electrode was useful for the elimination of ascorbic acid interference of the glucose sensor.

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A Biotin-avidin Labeled Enzyme Immunoassay for the Quantitation of Serum TSH Using Protein-layered Solid Phase

  • Choi, Myung-Ja;Song, Eun-Young;Chung, Tai-Wha
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.231-235
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    • 1998
  • A sensitive enzyme immunoassay for serum TSH has been developed utilizing the tight binding between biotin and avidin, and three layered protein polystyrene beads as solid phase. To increase binding capacity of TSH and sensitivity of the assay, the polystyrene beads were coated sequentially with mouse immunoglobulin as first layer, rabbit antimouse immunoglobulin as second layer and monoclonal anti-TSH as third layer. A serum sample was incubated simultaneously with a monoclonal anti-TSH immobilized polystyrene beads and a second monoclonal anti-TSH covalently attached to biotin. After washing, the antibody bound serum TSH-anti-TSH-biotin complex is reacted with horseradish peroxidase (HRP)-labeled avidin. Following second wash, the bound HRP activity was measured calorimetrically. Reproducible results were obtained within 4 hours for serum TSH in the range between $0{\mu}\textrm{IU}$ml and ${50}{\mu}\textrm{IU}$ml with detection limit of $0.1{\mu}\textrm{IU}$ per test.

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Analysis of Avidin-biotinylated Liposome Layers on Au Electrode by Quartz Crystal Analyzer (수정진동자를 이용한 Au 표면에서 avidin-biotin 결합 리포좀 막의 형성구조 분석)

  • Song, Seong-Hun;Cho, Hong-Sig;Park, Jong-Won;Kim, Kwang;Nakamura, Chikashi;Yang, Qing;Miyake, Jun;Chang, Sang-Mok
    • KSBB Journal
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    • v.15 no.5
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    • pp.497-500
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    • 2000
  • Liposomes and proteoliposomes, artificial membranes, can interact with many solutes, such as drugs, peptides and proteins. The immobilization of (prot대)liposomes as supramolecular aggregates on gold surfaces have potential applications in nano and biosensor technology. We demonstrated a quartz crystal analyzer (QCA) based method to monitor the construction of multi-layers of unilamellar liposomes based on avidin-biotin binding on gold surfaces using a quartz crystal microbalance (QCM). Thus, the QCA provides an on line and efficient method of detecting the construction of protein membranes, which has applications in biosensing systems.

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Detection of Avidin Based on Rugate-structured Porous Silicon Interferometer

  • Koh, Young-Dae;Kim, Sung-Jin;Park, Jae-Hyun;Park, Cheol-Young;Cho, Sung-Dong;Woo, Hee-Gweon;Ko, Young-Chun;Sohn, Hong-Lae
    • Bulletin of the Korean Chemical Society
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    • v.28 no.11
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    • pp.2083-2088
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    • 2007
  • Biosensor based on rugate PSi interferometer for the detection of avidin has been described. Rugate PSi fabricated by applying a computer-generated pseudo-sinusoidal current waveform has been prepared for the application as a label-free biosensor based on porous silicon interferometer. The fabrication, optical characterization, and surface derivatization of a rugate PSi has been described. The method to fabricate biotinderivatized rugate PSi has been investigated. The surface and cross sectional morphology of rugate PSi are obtained with SEM. FT-IR spectroscopy is used to characterize the oxidation and functionalization reaction of rugate PSi sample. Binding of the avidin into the biotin-derivatized rugate PSi induces a change in refractive index. A red-shift of reflectivity by 18 nm in the reflectivity spectrum is observed, when the biotin-modified rugate PSi was exposed to a flow of avidin.

Design, Synthesis and Preliminary Biological Evaluation of a Biotin-S-S-Phosphine Reagent

  • Kang, Dong W.;Kim, Eun J.
    • Bulletin of the Korean Chemical Society
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    • v.35 no.2
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    • pp.383-391
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    • 2014
  • Biotin-S-S-Phosphine was designed and synthesized as a potential tool for a proteomic study of O-GlcNAcmodified proteins. This reagent features a disulfide linker between a triarylphosphine moiety, which allows selective conjugation to azide-containing proteins, and a biotin moiety that can allow easy isolation through its strong affinity toward avidin-coated solid beads. The disulfide linkage within this reagent can allow the easy release of the bound molecules of interest, which is difficult to achieve when a biotin:avidin pair is used alone, by reducing the disulfide bond of the reagent with DTT. Preliminary in vitro biological assays with azidelabeled and unlabeled cell lysates and a pure protein Nup62 showed that the Biotin-S-S-Phosphine reagent is highly reactive toward the free thiol groups of proteins. When a molecular tool with a disulfide linker is applied to the enrichment of the molecules of interest from other species, it is important to block the free-thiols of the sample using exhaustive alkylation prior to the Staudinger ligation reactions to restore the bioorthogonal nature of this reaction.

Avidin-biotin complex for immunohistochemical diagnosis of Aujeszky's disease and hog cholera (Avidin-biotin 복합체를 이용한 오제스키병과 돼지콜레라의 면역조직화학적 감별진단)

  • Kim, Soon-bok;Sur, Jung-hyang;Moon, Un-gyeong
    • Korean Journal of Veterinary Research
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    • v.30 no.4
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    • pp.435-440
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    • 1990
  • Ten pigs infected with Aujeszky's disease virus (ADY) or hog cholera virus(HCV) were tested for the detection of virus antigens in frozens or paraffin-embedded sections by avidin-biotin-peroxidase complex(ABC) method. Tonsils, spleens, cerebra and buffy coats were examined for the immunohistochemical test. Where ADV antigen was detected by ABC, a dark brown deposit occurred in both the nucleus and the cytoplasm of lymphocytes and macrophages, however, HCV antigen was demonstrated in the cytoplasm of the infected cells. ADV-positive cells were most frequently detected in tonsils and cerebra, whereas, HCV -positive cells were frequently observed in spleens. And buffy coat were also good for both virus detection. The results suggested that ABC method is considered as an excellent and reliable tool for confirmative diagnosis of these viral diseases.

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High-Contrast Imaging of Biomolecular Interactions Using Liquid Crystals Supported on Roller Printed Protein Surfaces

  • Park, Min-Kyung;Jang, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.33 no.10
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    • pp.3269-3273
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    • 2012
  • In this study, we report a new method for the high contrast imaging of biomolecular interactions at roller printed protein surfaces using thermotropic liquid crystals (LCs). Avidin was roller printed and covalently immobilized onto the obliquely deposited gold surface that was decorated with carboxylic acid-terminated self-assembled monolayers (SAMs). The optical response of LCs on the roller printed film of avidin contrasted sharply with that on the obliquely deposited gold surface. The binding of biotin-peroxidase to the roller printed avidin was then investigated on the obliquely deposited gold substrate. LCs exhibited a non-uniform and random orientation on the roller printed area decorated with the complex of avidin and biotin-peroxidase, while LCs displayed a uniform and planar orientation on the area without roller printed proteins. The orientational transition of LCs from uniform to non-uniform state was triggered by the erasion of nanometer-scale topographies on the roller printed surface after the binding of biotin-peroxidase to the surface-immobilized avidin. The specific binding events of protein-receptor interactions were also confirmed by atomic force microscopy and ellipsometry. These results demonstrate that the roller printing of proteins on obliquely deposited gold substrates could provide a high contrast signal for imaging biomolecular interactions using LC-based sensors.