• Title/Summary/Keyword: Avidin

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생물 공학 의약품의 품질관리에 관한 연구(비방사능 물질 표지법을 이용한 숙주유래 DNA의 검출법 개발)

  • 용군호;민홍기;김창민;오호정;한강현;최규실
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.59-59
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    • 1993
  • 비방사능물질인 Biotin을 DNA probe에 표지하여 nonisotopic hybridization 방법을 사용하여 감도를 높임으로써, 손쉽게 생물공학 의약품의 품질관리에 사용되도록 하였다. Bethesda Research Laboratories(BRL) 회사 제품인 biotinylated probes-avidin alkaline phosphatase를 이용한 chemiluminescene detection방법으로 행하여 λ phage DNA, yeast DNA, E.coil DNA의 한계 검출 농도를 알아내고, 생물 공학 제품에 적용하였다. Dot blot hybridization 방법으로 행하여 λ phage DNA는 0.1pg, Yeast DHA는 4.5pg, E. coli DNA는 8.9pg까지 검출되었고, 기존 생물공학 제품에서는 숙주 유래 DNA가 전혀 검출되지 않았다.

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Genetically engineered brain drug delivery vector through the blood-brain barrier

  • Seo, Kyung-Hee;Kang, Young-Sook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.192-192
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    • 1998
  • The blood - brain barrier (BBB) expresses high concentrations of transferrin receptor, and it was revealed that anti-transferrin receptor mouse monoclonal antibody (OX26) undergoes transcytosis through the BBB. This property allows the OX26 to serve as a brain drug delivery vector. In an attempt to produce broadly useful targeting agents, genetic engineering and expression techniques have been used to produce antibody-avidin (AV) fusion protein (OX26 IgG3C$\_$H/3-AV). In the present study we estimated the BBB permeability and stability of genetically engineered vector.

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Isolation and immunohistochemical diagnosis of Haemophilus parasuis from naturally occurring polyserositis in pigs (자연발생한 돼지 다발성 장막염 예로부터 Haemophilus parasuis의 분리와 면역조직화학적 진단)

  • Bae, You-chan;Kang, Mun-il;Hwang, Eui-kyong;Sohn, Hyun-joo;Choi, Sang-ho
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.843-852
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    • 1998
  • From Jan. 1996 to Oct. 1997, 29 pigs with 40-70 days old showing dyspnea inappetite and polyserositis were collected and carried out necropsy, bacterial culture, histopathology, avidin biotin complex(ABC) stain, fluorescent antibody(FA) test, and electron microscopy. In the study, 4 strains from 3 pigs were isolated from meninges, pleura and synovial fluid and also were identified as Haemophilus parasuis serovar 5. Main histopathological lesions of 29 pigs with polyserositis were frequently composed of fibrinous peritonitis(27), pleurisy(22), interstitial pneumonia(21), fibrinous epicarditis(20), fibrinopurulent meningitis(8) and synovitis(4). By ABC stain, 11/29(37.9%) pigs with polyserositis were confirmed to have H parasuis serovar 5 in the cytoplasm of macrophages and neutrophils in cerebral meninges, epicardium, pleura surface of lung or serosa of spleen. ABC stain(20.8~40.0%) to detect H parasuis serovar 5 in tissues was more sensitive than bacterial culture(10.3%), but less sensitive than FA test(62.5%) using frozen tissues even though the result of 8 cases. By electron microscopy, a bacterium was also detected in the cytoplasm of macrophages in purulent exudate of cerebral meninges. Consequently, we confirmed that H parasuis serovar 5 has been involving to cause pigs with polyserositis and can be detected by FA and ABC stain as reliable diagnostic tools.

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ANALYSIS OF FLUIDIC BEAD CUBE EMBEDDED PORTABLE CMOS SENSING SYSTEM FOR IMMUNO REACTION MONITORING (유체소자가 집적화된 면역검사용 휴대용 CMOS 바이오칩의 분석)

  • Jeong, Yong-Won;Park, Se-Wan;Kim, Jin-Seok;Kim, Hyeon-Cheol;Chun, Kuk-Jin
    • Proceedings of the IEEK Conference
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    • 2005.11a
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    • pp.755-758
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    • 2005
  • This paper describes the novel immunoassay sensing system for a portable clinical diagnosis system. It consists of a bead cage reactor and a CMOS integrated biosensor. It showed the simple and easy antibody coating method on beads by flow-through avidin biotin complex technology in a microfluidic device. It showed just 90 nL sample consumption and good result for the application of alpha feto protein. The bead cage reactor has the role of the antibody coating, antigen binding and enzyme linking for the electrochemical sensing method. The CMOS biosensor consists of ISFET (ion selective field effect transistor) biosensor and temperature sensor for detecting pH that is the byproduct of enzyme reaction. The sensitivity is 8 $kHz/^{\circ}C$ in a temperature sensor and 33 mV/pH in a pH sensor. After filling the 15 um polystyrene beads in bead cage, antibody flowed and reacted to beads. Subsequently, the biotinylated antigen flowed and bound to the antibody and GOD (glucose oxidase)-avidin conjugate flowed and reacted to the biotin of the biotinylated antigen. After this reaction process, glucose solution flowed and reacted to the GOD on beads. The hydrogen was generated by glucose-GOD reaction. And it was detected by the pH sensor.

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The selection of basic platform for improving the sensitivity of neutravidin rapid detection kit (뉴트라비딘 검출용 간이 진단키트의 성능향상을 위한 기본 플랫폼 선정)

  • Choi, Sunmi;Kim, Giyoung;Om, Aeson;Moon, Jihea;Park, Saetbyeol;Lee, Sangdae;Kim, Hyuk Joo
    • Korean Journal of Agricultural Science
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    • v.39 no.4
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    • pp.613-618
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    • 2012
  • This study was performed to optimize the basic platform of a lateral flow immunoassay. Improvement of the limit of detection (LOD) was evaluated according to the width of a nitrocellulose membrane with varying concentrations of analyte. The analyte, neutravidin was detected based on the avidin-biotin interaction. The antibody-Au nanoparticle conjugation was mostly stabled in a PBS buffer of pH 7.3. The optimal widths of a nitrocellulose membrane were 4 and 6 mm considering the sample flow rate and signal strength of the test line on the membrane. The LOD of neutravidin was 0.001 mg/ml in the optimum conditions.

Significance of a Highly Specific and Sensitive Enzyme Linked Immunosorbent Assay on Evaluation of Environmental Toxicant-Mediated Allergic Responses

  • Kim, Hyoung-Ah;Yong Heo
    • Toxicological Research
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    • v.17
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    • pp.197-199
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    • 2001
  • Enhancement of antigen-specific IgE is a hallmark of allergic hyperresponsiveness, therefore it is necessary to adopt or develop a highly sensitive and specific assay for determination of allergen-specific IgE levels in vivo. In this presentation, we introduce an ELISA (enzyme linked immunosorbent assay) system developed to measure the levels of chicken egg ovalbumin (OVA)-specific IgE in serum. The ELISA method uses a commercially available purified rat anti-mouse IgE as a capture Ab and biotinylated OVA as a detection reagent. Avidin-peroxidase with its substrate is used for color development resulting in optical density measurement at 405 nm. The ELISA system produces a highly sensitive dose-response relation-ship between optical density levels and the dilution titer of the OVA-IgE standard serum but no cross-reaction with unrelated IgE or IgG. It is believed that the system is an Efficient tool to delineate an adjuvant effect of environmental pollutants on development of asthmatic and atopic responses.

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Primary Cellular Study of Phagocytosis for Alzheimer Disease Diagnosis (알츠하이머 조기 진단을 위한 변형된 대식세포의 기초적 연구)

  • Cho, Jung-Min;Chae, Cheol-Joo;Kang, Jae-Min;Kim, Kwan-Su;Song, Ki-Bong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2010.06a
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    • pp.280-280
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    • 2010
  • Alzheimer disease is a progressive neurodegenerative disease of the aged, characterized by memory loss and dementia. For diagnosis of Alzheimer disease we have simply modified macrophage with amyloid beta bonded with different molecules. Modified Macrophage was observed with microscope for co-localization of amyloid beta molecule. For this experiment we used fluoroscene labeling substances. The macrophage was modified also with cell staining method. For cell staining method was used avidin-biotin reaction principles. All experiments were carried out on poly-L-lysine coated and sterilized glass substrates. In the presentation we will show the further investigations and applications with modified macrophage.

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AN IMMUNOHISTOCHEMICAL LOCALIZATION OF TENASCIN IN PERIODONTAL POCKET TISSUES (치주낭 조직내 tenascin의 분포에 관한 면역조직화학적 연구)

  • Han, Kyung-Yoon;Lee, Kang-Jin
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.607-617
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    • 1994
  • To determine the effect of tenascin on forming periodontal pocket and pseudopocket, the ginival tissues were surgically obtained from the patients with adult periodontitis(10) and non-inflammatory phenytoin-associated gingival hyperplasia(5). The excised tissue specimens were fixed in neutral formalin for $6{\sim}24$ hours, embedded with paraffin, sectioned at 4-6m in thickness, mounted on glass slides coated with 3-aminopropyltriethoxysilane(Sigma Chemical Co., St. Louis, MO, U.SA.) and immunohistochemically processed by Avidin-Biotin peroxidase complex method for the localization of tenascin, using monoclonal mouse anti-human tenascin antiboday(Chemicon-International Inc., Temecula, CA, U.S.A., 1: 5,000) as the primary antibody. Regardless of periodontal pocket and pseudopocket, tenascin was localized along the connective tissue subjacent to basement membrane of gingival epithelium, and strong positive reactivity was obviously noted in the papillary projections of gingival connective tissue. The results suggest that tenascin may affect the development of papillary projections and the proliferation of epithelial cells.

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MELANOTIC NEUROECTODERMAL TUMOR OF INFANCY ; A CASE REPORT (유아기 흑색 신경외배엽성 종양의 치험례)

  • Kim, Il-Kyu;Ha, Soo-Yong;Lee, Seong-Jun;Chu, Young-Chae
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.13 no.4
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    • pp.436-443
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    • 1991
  • A case of melanotic neuroectodermal tumor of infancy(MNTI) in 5 month old girl is presented with review of the literature. The review of literature indicated that there have been 189 reported cases of this lesion and there were 9 cases of malignant tumor for a rate of 4.8%. This rate of malignancy is very high in view of the fact that the tumor has been described as benign. And immunohistochemical study using avidin-biotin conjugate method demonstrated that this tumor had been originated from neural crest.

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Preparation and Atomic Force Microscopy (AFM) Characterization of DNA Scaffolds as a Template for Protein Immobilization

  • Kim, Hyeran;Lee, Hyun Uk;Lee, Jouhahn
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.411.2-411.2
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    • 2014
  • The design of DNA nanostructures is of fundamental importance, the intrinsic value of DNA as a building-block material lies in its ability to organize other bio-molecules with nanometer-scale spacing. Here, we report the fabrication of DNA scaffolds with nano-pores (<10 nm size) that formed easily without the use of additives (i.e., avidin, biotin, polyamine, or inorganic materials) into large-scale structures by assembling DNA molecules at near room temperature ($30^{\circ}C$) and low pH (~5.5). Protein immobilization results also confirmed that a fibronectin (FN) proteins/large scale DNA scaffolds/aminopropylytriethoxysilane (APS)/SiO2/Si substrate with high sensitivity formed in a well-defined manner. The DNA scaffolds can be applied for use with DNA-based biochips, biophysics, and cell biology.

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