• Title/Summary/Keyword: Avidin/Biotin

Search Result 102, Processing Time 0.023 seconds

수정진동자를 이용한 Au 표면에서 avidin-biotin 결합 리포좀 막의 구조 분석

  • Park, Jong-Won;Han, Seong-Ung;Gwon, Jeong-Hun;Park, Jin-Yeong;Jo, Hong-Sik;Lee, Haeng-Ja;Jang, Sang-Mok
    • 한국생물공학회:학술대회논문집
    • /
    • 2000.04a
    • /
    • pp.546-549
    • /
    • 2000
  • Liposomes and proteoliposomes, artificial membranes, can interact with many solutes, such as drugs, peptides and proteins. Immobilization of (proteo)liposomes as supramolecular aggregates on gold surfaces have potential applications in nanotechnology and biosensors. We demonstrate a quartz crystal analyzer (QCA) method to monitor the construction of multi layers of unilamellar liposomes based on avidin-biotin binding on gold surface using quartz crystal microbalance(QCM). Thus, QCA provides an on line and efficient method to detect the protein membranes construction and have applications to biosensing system.

  • PDF

Potentiometric Homogeneous Enzyme-Linked Binding Assays for Riboflavin and Riboflavin Binding Protein

  • 김진목;김혜진;김미정;이동주;한상현;차근식
    • Bulletin of the Korean Chemical Society
    • /
    • v.17 no.11
    • /
    • pp.1018-1022
    • /
    • 1996
  • Adenosine deaminase (ADA) has been utilized as the label in devising a potentiometric homogeneous assay for riboflavin and riboflavin binding protein (RBP). The proposed homogeneous assay method employs an ADA-biotin conjugate as the signal generator and an avidin-riboflavin conjugate as the signal modulator in the solution phase. The catalytic activity of the ADA-biotin conjugate is inhibited in the presence of an excess amount of the avidin-riboflavin conjugate, and the observed inhibition is reversed in an amount proportional to the concentration of RBP added. When the analyte riboflavin is added to this mixture of ADA-biotin, avidin-riboflavin and RBP, the activity of the enzyme conjugate is re-inhibited in an amount proportional to the concentration of riboflavin. Since the enzyme label used in this system is ADA, an ammonia-producing enzyme, a potentiometric rather than photometric detection scheme is used to monitor the enzymatic activity in the assay.

Biodistribution Study of $^{99m}Tc$-Labeled Succinic Acid-Conjugated Low pI Avidin (낮은 동전점을 갖는 $^{99m}Tc$ 표지 Succinic Acid 결합 Avidin의 생체내분포에 관한 연구)

  • Jeong, Jae-Min;Paik, Chang-H.
    • The Korean Journal of Nuclear Medicine
    • /
    • v.27 no.2
    • /
    • pp.285-292
    • /
    • 1993
  • Avidin과 biotin의 높은 결합력을 이용하여 종양 영상을 개선하는 방법이 많이 연구되고 있다. 본 실험에서는 이러한 목적으로 쓰기 위하여 적당한 $^{99m}Tc$ 표지 avidin을 제조하였다. Avidin을 표지하기 위하여 우선 $^{99m}Tc$과 안정한 킬레이트를 형성할 수 있는 benzoylmercaptoacetyltriglycine (Bz-MAG3)과 biocytin을 화학적으로 결합시킨 Bz-MAG3-biocytin을 합성하였다. 이 화합물을 Tc-99m으로 표지시켜 avidin 또는 streptavidin을 1:1로 섞어 줌으로서 Tc-99m으로 표지된 avidin과 streptavidin을 제조하였다. 이들의 생쥐 생체내 분포를 조사한 결과 avidin의 경우 높은 간(56.6%, 10min)과 신장(28.5%, 10min) 축적을 보였고 streptavidin의 경우 높은 신장 축적 (28.9%, 21hr)을 보였다. Avidin의 높은 정상 조직 축적을 줄이기 위하여 succinic acid를 결합시켜 등전점(pI)을 낮춘 다음 같은 실험을 하여 본 결과 신장 축적율은 pI가 $7.0{\sim}9.3,\;5.5{\sim}6.2,\;4.0{\sim}4.8$로 낮아졌을 경우 19.0%, 3.1%, 1.7%로 각각 떨어졌지만 간에의 축적은 pI 변화에 따른 상관성을 찾아 볼 수가 없었다. 체내 제거율을 측정하여 본 결과 pI를 변화시킨 avidin과 변화시키지 않은 avdin들은 반감기가 13.5에서 16.0시간 사이로 큰 차이점을 보이지 않았는데 streptavidin은 반감기 61.5시간 정도로 느리게 제거된다는 것을 알았다. 이 실험의 결과 1. Avidin을 $^{99m}Tc$-MAG3-biocytin으로 안정하게 표지할 수 있었고, 2. pI가 낮아진 avidin은 신장에의 축적율이 크게 감소되었으며, 3. $^{99m}Tc$으로 표지된 avidin과 streptavidin은 먼저 간으로 흡수된 후 대사된 다음 신장으로 배설된다는 사실을 알았다.

  • PDF

Patterning Biological Molecules onto Poly(amidoamine) Dendrimer on Gold and Glass

  • Hong, Mi-Young;Lee, Do-Hoon;Yoon, Hyun C.;Kim, Hak-Sung
    • Bulletin of the Korean Chemical Society
    • /
    • v.24 no.8
    • /
    • pp.1197-1202
    • /
    • 2003
  • Patterning of biological molecules was attempted on both gold and glass using fourth generation (G4) poly(amidoamine) (PAMAM) dendrimer as an interfacing layer between solid surfaces and biomolecules. As for the patterning of avidin and anti-biotin antibody on gold, PAMAM dendrimers representing amine functionalities were firstly printed onto the 11-mercaptoundecanoic acid SAM by microcontact printing, followed by biotinylation, and reacted with fluorescence-labeled avidin or anti-biotin antibody. Fluorescence microscopic analysis revealed that the patterns of avidin and anti-biotin antibody were well constructed with the resolution of < 2 ㎛. The PAMAM dendrimers were also printed onto aldehyde-activated slide glass and reacted directly with anti-BSA antibodies, which had been oxidized with sodium periodate. As a result, distinct patterns of the anti-BSA antibodies were also obtained with a comparable edge resolution to that of avidin patterns on gold. These results clearly show that PAMAM dendrimers can be adopted as an interfacing layer for the patterning of biological molecules on solid surfaces with micrometer resolution.

Fabrication and Characterization of Pyrolyzed Carbon for Use as an Electrode Material in Electrochemical Biosensor (전기화학 바이오센서의 전극물질로 응용을 위한 열분해 탄소의 제작 및 특성 연구)

  • Lee, Jung-A.;Hwang, Seong-Pil;Kwak, Ju-Hyoun;Park, Se-Il;Lee, Seung-Seob;Lee, Kwang-Cheol
    • Transactions of the Korean Society of Mechanical Engineers A
    • /
    • v.31 no.10
    • /
    • pp.986-992
    • /
    • 2007
  • This paper presents the fabrication and characterization of carbon films pyrolyzed with various photoresists for bioMEMS applications. To verify the usefulness of pyrolyzed carbon films as an electrode material in an electrochemical biosensor developed by the authors, interactions between avidin and biotin on the pyrolyzed carbon film were studied via electrochemical impedance spectroscopy based on electrostatic interactions between avidin and negatively-charged ferricyanide. The pyrolyzed carbon films were characterized using a surface profiler, a precision semiconductor parameter analyzer, a nanoindentor, scanning electron microscopy, and atomic force microscopy. Amine conjugated biotin was immobilized on the electrode using EDC/NHS as crosslinkers after $O_2$ plasma treatment to enhance functional groups on the carbon electrode pyrolyzed at $1000^{\circ}C$ with AZ9260. The detection of avidin binding with different concentrations in a range of 0.75 nM to $7.5\;{\mu}M$ to the pyrolyzed carbon electrode modified with biotin was carried out by measuring the electrochemical impedance change. The results show that avidin binds to the biotin on the electrode not by non-specific interaction but by specific interaction, and that EIS successfully detects this binding event. Pyrolyzed carbon films are a promising material for miniaturization, integration, and low-cost fabrication in electrochemical biosensors.

Enzyme-Linked Immuno-Sorbent Assay for Bovine Caseins (우 Casein의 면역효소분석법)

  • 염행철
    • Korean Journal of Animal Reproduction
    • /
    • v.16 no.2
    • /
    • pp.87-102
    • /
    • 1992
  • A rapid, sensitive, and specific enzyme-linked immuno-sorbent assay (ELISA) for bovine casein was developed. Biotinylated casein and peroxidase-conjugated avidin were used in the assay with antibody separated from yolks of immunized hens. Caseins were biotinylated with sulfo-N-hydroxy succinimido biotin and peroxi-dase-conjugated avidin bound the biotinylated casein which became bound to immobilized anti-body on a microplate. The antibodies were specific for bovine $\alpha$- and $\beta$-caseins, and their cross-reactivities with whey proteins, IgG, and serum albumin from bovine were not detectable by ELISA and Western blot. Various sensitivities ranging from 2ng/ml to 20${\mu}\textrm{g}$/ml of casein were achieved, and were controlled by adding vanous concentrations of the biotinylated casein. Parallelism was observed between standard and sample curves. The coefficients of variation of intra-assays and inter-assays from the most sensitive assay were 5.5 and 5.7%, respectively, at the 50% displacement. Casein contents of peripaturient milk samples showed that casein secretion rapidly increased 3d prepartum.

  • PDF

Congenital transmission of Theileria sergenti in cattle verified by immunohistochemistry (소 Theileria sergenti의 태반감염에 대한 면역세포화학적 증명)

  • Baek, Byeong-kirl;Kim, Jin-ho;Onuma, Misao
    • Korean Journal of Veterinary Research
    • /
    • v.37 no.4
    • /
    • pp.825-829
    • /
    • 1997
  • The spleen and placenta from the aborted fetuses as well as lymphnodes and placenta of the corresponding dams naturally infected with T sergenti were used to localize the parasite antigens by immunohistochemical staining for the possible congenital transmission of theileriosis. Parasite-specific antigens were detected immunohistochemically by incubating the sections with specific monoclonal antibody prepared against 34KD surface antigen of T sergenti and visualized via the avidin biotin complex(ABC) method. Specific T sergenti antigen was detected in the sections of formalin or acetone-fixed fetal spleens and placenta. Similar antigens were also demonstrated in lymphnodes and placentas of the corresponding dams. It is concluded that this technique will eventually play an important role in specialized diagnostic laboratories in the verification/evaluation of congenital infection with T sergenti.

  • PDF

Fabrication and Characterization of an OTFT-Based Biosensor Using a Biotinylated F8T2 Polymer

  • Lim, Sang-Chul;Yang, Yong-Suk;Kim, Seong-Hyun;Kim, Zin-Sig;Youn, Doo-Hyeb;Zyung, Tae-Hyoung;Kwon, Ji-Young;Hwang, Do-Hoon;Kim, Do-Jin
    • ETRI Journal
    • /
    • v.31 no.6
    • /
    • pp.647-652
    • /
    • 2009
  • Solution-processable organic semiconductors have been investigated not only for flexible and large-area electronics but also in the field of biotechnology. In this paper, we report the design and fabrication of biosensors based on completely organic thin-film transistors (OTFTs). The active material of the OTFTs is poly(9,9-dioctylfluorene-co-bithiophene) (F8T2) polymer functionalized with biotin hydrazide. The relationship between the chemoresistive change and the binding of avidin-biotin moieties in the polymer is observed in the output and on/off characteristics of the OTFTs. The exposure of the OTFTs to avidin causes a lowering of ID at $V_D$ = -40 V and $V_G$ = -40 V of nearly five orders of magnitude.

Homogeneous Enzyme-Linked Binding Assay Mediated by the Interaction of Avidin with Biotin: Mistletoe Lectin I Assay

  • Rhee Paeng, In-Suk;Lee, Eun-Ah;Kim, Hyun-Sook
    • Bulletin of the Korean Chemical Society
    • /
    • v.25 no.1
    • /
    • pp.115-118
    • /
    • 2004
  • We have examined the feasibility of using the specific interaction between mistletoe lectin I (ML I) and ${\beta}$-Dgalactose instead of the anti-ML I antibody in developing a homogeneous type competitive binding assay for ML I. We also have examined the feasibility of adapting the biotin/avidin mediated homogeneous assay for this system. Alkaline phosphatase (AKP) was employed as a single substrate enzyme label. The dose-response curve shows a detection range of 1-25 ${\mu}$g/mL and a linear response with a correlation coefficient of 0.99. To demonstrate the analytical utility of this method, 10 ${\mu}$g/mL of ML I was spiked into distilled water. The results show that the mean recovery was 10.03 ${\mu}$g/mL with an SD of 0.18. The difference between the spiked value and the mean recovery was 0.03 ${\mu}$g/mL, with a relative error of 0.3 and 1.6 % of RSD.