• Title/Summary/Keyword: streptavidin

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Detection of Protein Molecules by Electrical Current Response Using Two-Electrode Method

  • Lyu, Hong-Kun;Woo, Sung-Ho;Han, Yoon-Soo;Lee, Hee-Ho;Shin, Jang-Kyoo
    • Journal of Sensor Science and Technology
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    • v.20 no.2
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    • pp.90-95
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    • 2011
  • In order to protect human lives from disease, various biosensors having the potential to analyze a variety of biomolecules have been utilized. Biosensors constitute one of the most promising ways to monitor and detect various biomolecules corresponding to diseases. In this study, we demonstrate that the reaction of streptavidin molecules with biotin on a gold electrode can be detected using the twoelectrode method with a gold electrode and a platinum reference electrode. We also show the characteristics of the electrical current response. While detecting 2-${\mu}M$ streptavidin molecules dissolved in phosphate buffered saline(PBS) solution, we found that an analytical biosensor can operate on the principle of detecting an antigen-antibody reaction event of protein molecules using the two-electrode method. We think that the "potential step" method might be useful to detect the occurrence of any antigen-antibody reactions and can be combined with other devices or ICs such as BJTs, MOSFETs, and OP-amps for the detection of biomolecules of diseases.

Study on Bead-based Microbiochip and Analytical System for Protein Detection

  • Kim, Min-Soo;Chung, Woo-Jae;Cho, Su-Hyung;Park, Sung-Soo;Kim, Byung-Gee;Lee, Young-Sik;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2002.11a
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    • pp.60-63
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    • 2002
  • This paper presents bead-based microbiocihps to detect and separate target proteins. Micro beads coated with capture proteins were introduced into a microchamber, and target proteins flowing across the chamber were bound and concentrated. The chip was connected with an external fluid system. Bead surfaces were double-coated with photo-cleavable linkers and capture proteins. The proteins bound on the beads were photo-separated under UV irradiation, and excited to be measured in fluorescence. $38{\sim}50{\mu}m$ sized polystyrene beads were used. SOGs(silicon-on-glass) were used to fabricate the microchip having glasses bonded on both sides. 100 ${\mu}m$ thick silicon channel was formed through silicon deep RIE process. The upper glass cover had holed through to have inlets and outlets fabricated by powder-blastings. In this study, biotin and streptavidin were used as capture proteins and detection proteins, respectively. The protein mixtures of streptavidin, HSA(human serum albumin) and ovalbumin were applied for selective detection test.

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Salmonella species 검출용 DNA Probe 분석시스템 고안

  • Lee, Ung-Hui;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.711-712
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    • 2000
  • DNA probe assay comprising a microwell as' solid matrix for the immobilization of streptavidin (SA) and an oligonucleotide with covalently bound fluorecein as detection probe was developed. The insolubilized SA captured the biotinylated DNA product of polymerase chain reaction (PCR), and the product was denatured under a basic condition. The remaining single-stranded DNA on the solid surface was hybridized with the probe for signal generation that was performed based on enzyme-linked immuno -reactions.

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Immunohistochemical detection of infectious hematopoietic necrosis virus antigens in cell cultures (배양세포에서 전염성조혈장기괴사증 바이러스항원의 면역조직화학적 검출)

  • 문운경;이민권;진영배;김순복
    • Korean Journal of Veterinary Service
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    • v.25 no.3
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    • pp.295-297
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    • 2002
  • This experiment was done to set up the immunohistochemical detection method for infectious hematopoietic necrosis virus(IHNV) antigens in the monolayers of CHSE-214 cell cultures inoculated with IHNV. Specific identification of IHNV antigens was detected in the cytoplasms of infected cells by the use of monoclonal antibodies to glycoproteins. The specific positive signal was observed as a distinct red color. The result showed that streptavidin alkaline phosphatase immunohistochemistry specifically identified IHNV antigens in infected cultured cells.

Microcontact Printing of Biotin for Selective Immobilization of Streptavidin-fused Proteins and SPR Analysis

  • Lee, Sang-Yup;Park, Jong-Pil;Lee, Seok-Jae;Park, Tae-Jung;Lee, Kyung-Bok;Park, Insung S.;Kim, Min-Gon;Chung, Bong-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.137-142
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    • 2004
  • In this study, a simple procedure is described for patterning biotin on a glass substrate and then selectively immobilizing proteins of interest onto the biotin-patterned surface. Microcontact printing (CP) was used to generate the micropattern of biotin and to demonstrate the selective immobilization of proteins by using enhanced green fluorescent protein (EGFP) as a model protein, of which the C-terminus was fused to a core streptavidin (cSA) gene of Streptomyces avidinii. Confocal fluorescence microscopy was used to visualize the pattern of the immobilized protein (EGFP-cSA), and surface plasmon resonance was used to characterize biological activity of the immobilized EGFP-cSA. The results suggest that this strategy, which consists of a combination of $\mu$CP and cSA-fused proteins. is an effective way for fabricating biologically active substrates that are suitable for a wide variety of applications. one such being the use in protein-protein assays.

Electrochemical Detection of $17{\beta}-estradiol$ by using DNA Aptamer Immobilized Nanowell Gold Electrodes

  • Kim, Yeon-Seok;Jung, Ho-Sup;Lee, Hea-Yeon;Kawai, Tomoji;Gu, Man-Bock
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.88-92
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    • 2005
  • Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

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Characterization of T7 RNA Polymerase Transcription Elongation Complex in Sequence-specific Transcription Termination (염기서열 특이적 전사종결부위에서 T7 RNA 중합효소 전사연장복합체 특성에 관한 연구)

  • Shin, Ji-Young;Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.14 no.1
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    • pp.39-50
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    • 2004
  • T7 RNA polymerase is a single subunit RNA polymerase able to accomplish whole transcription process without auxiliary factors. In order to study transcription elongation mechanism of phage T7 RNA polymerse, stepwise walking of RNA polymerase was established by immobilizing biotinylated DNA template with streptavidin bead, series of active and stable elongation complexes were obtained, Transcripts were radio isotope labeled at the 16thm 17th and 18th nucleotide residues so stable elongation transcription complex of T7 RNA polymerase containing 22-40 nucleotide residues could be identified. We identified the positions of stablely formed transcription elongation complexes of termination site in intrinsic hairpin-independent PTH terminator sequence through the established stepwise walking of wild-type of mutant R173C T7 RNA polymerases. The results suggest that stable elongation transcription complexes were at the site of passing PTH terminator signal by mutant R173C RNA polymerase.

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Development of Human Papillomavirus DNA Array by Using Lateral Flow Membrane Assay (Lateral Flow Membrane를 이용한 인유두종 바이러스 DNA Array의 개발)

  • Kim, Ki-Whang;Lee, Hyung-Ku;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.346-351
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    • 2008
  • This study develops DNA array which can detect specific sequence of human papilomavirus (HPV) by using lateral flow membrane assay which is usually used for point of care test including pregnant diagnosis. Principle of HPV DNA array is as follow; fixing DNA probe which is peculiar to HPV type 6, 11, 16, 18, 31, 45 on a surface of lateral flow membrane and inducing hybridization response between probe and HPV PCR products which is obtained by using biotin-labeled MY09/l1 primers. And then, we can see the result of DNA hybridization that streptavidin labelled colloidal gold is responded with hybrid biotin. Lateral flow membrane array developed in this study confirms major HPV type economically and conveniently compared with existing HPV DNA chip method.

Design and fabrication of a highly sensitive microcalorimetric biosensor by bulk micromachining (벌크 마이크로 머시닝을 이용한 고감도 미세 칼로리미터의 설계 및 제작)

  • Yoon, S.I.;Kim, J.H.;Kwak, B.S.;Kim, Y.J.;Jung, H.I.
    • Journal of Sensor Science and Technology
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    • v.15 no.3
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    • pp.164-167
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    • 2006
  • Calorimeter is one of widely used biosensors. Conventional or existing calorimeters are realized directly on a silicon wafer which has very high thermal conductivity. It results in decreasing temperature difference between junctions and it makes a sensitivity of calorimeter to be decreased. In this study, the microcalorimeter was made by using MEMS(Micro Electro Mechanical Systems)-technology and hot junctions of the microcalorimeter are released from a silicon substrate to reduce loss of generated heat by reactions between biomolecules. Sensitivity of the released microcalorimeter was 18 mV/M which is 1.5 times higher than another calorimeters on silicon substrate by reactions between biotin and streptavidin.