• 분석과학
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• 제18권6호
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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.

• Archives of Pharmacal Research
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• 제21권3호
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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.

• 대한수의학회지
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• 제30권4호
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• pp.435-440
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• 1990

오제스키병바이러스 또는 돼지콜레라바이러스에 인공 또는 자연감염된 돼지 10두를 실험동물로 공시하였으며, 감염돈의 편도선, 비장, 대뇌 및 연층(buffy coat)의 냉동 및 파라핀절편에서, avidin-biotin-peroxidase complex(ABC)를 이용하여 이들 바이러스를 면역조직화학적으로 검출하였다. 오제스키병바이러스 항원은 임파구와 대식세포의 핵내 또는 세포질에서 검출되었으며, 돼지콜레라바이러스 항원은 이들 세포의 세포질에서 검출되었고, ABC법은 양성반응 부위에서 갈색의 색소 침착을 일으켰다. 오제스키병바이러스 양성세포는 편도선과 대뇌에서 가장 빈번히 검출되었음에 반해 돼지콜레라바이러스는 비장에서 가장 빈번하였다. 그리고 연충에서도 이들 두가지 바이러스항원이 모두 검출되었다. ABC법은 이을 바이러스의 면역조직화학적 검출에 있어 특이성이 높고 배경의 비특이염색성이 낮아, 바이러스 분리동정을 거치지않고 이들 질병을 확진할 수 있는 진단수단으로 활용될 수 있을 것으로 생각된다.

• 대한수의학회지
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• 제37권4호
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• pp.825-829
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• 1997

소 theileriosis의 태반감염 사실을 입증하기 위하여 Theileria sergenti에 자연감염된 암소의 비장, 태반 그리고 유산된 태자의 비장과 태반조직으로부터 T sergenti를 면역화학적으로 검색 입증하고자 T sergenti 표피의 34KD 항원의 단크론 항체를 활용하여 avidin biotin complex 방법으로 면역화학적으로 염색하였던 바, 이들 formalin 고정 조직표본에서 T sergenti의 특이 항원성 물질을 관찰함으로써 태반감염을 증명할 수 있었다.

• Bulletin of the Korean Chemical Society
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• 제33권10호
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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.

• 대한수의학회지
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• 제33권1호
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• pp.119-123
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• 1993

The present study was intended to use the avidin-biotin-peroxidase-antiperoxidase complex (ABPAP) method for the identification of Mycobacterium bovis in the tissue sections of infected cattle. Antibodies and linksera for ABPAP procedure used in incubated order were rabbit anti-Mycobacterium polyvalent antibodies, goat anti-rabbit IgG, rabbit peroxidase-antiperoxidase complex, biotinyl-horse anti-rabbit IgG, and avidin-biotin-peroxidase complex. Where the bacterial antigen was localized by ABPAP, a dark brown deposit occurred in the cytoplasms of macrophages and Langerhans' giant cells of the granulomatous lesions. The method approved to be highly specific for the identification of the bacteria and allowed a precise localization of the bacterial antigen in infected cells.

• 대한수의학회지
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• 제33권2호
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• pp.255-261
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• 1993

For a immunohistochemical diagnosis of the frozen and paraffin-embedded tissues against rabies virus, mice were intracerebrally inoculated with challege virus standard(CVS) rabies virus and then were used to detect the rabies viral antigen by the immunoperoxidase(IP) and the avidin-biotin complex(ABC) method. In this study, the results confirmed that ABC and IP methods, although the former showed more specific and sensitive than the latter, were reliable and effective for the demonstration of rabies virus in both frozen and paraffin-embedded brain tissues prepared from rabies-infected mice. Additionally, IP technique using the monoclonal antibody against rabies virus could be recommended as a standard diagnostic tool instead of the present immunofluorescent method for the local veterinary services in Korea.

• 대한의생명과학회지
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• 제2권2호
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• pp.275-282
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• 1996

저자들은 마우스를 실험모델로 하여 간흡충의 항원을 투여 했을 때 비장조직에 대한 CD3, CD4 및 CD8 모노클로날 항체의 반응 여부를 알아보고자 하였다. 즉, 간흡충에 대한 세포면역학적인 특성을 규명고자 하였으며 특히 비장 조직에 대한 phenotype을 관찰한 결과 다음과 같은 결과를 얻었다. 간흡충의 조항원을 면역증강제와 함께 복강 투여한 다음 일정 기간 후에 비장조직을 Avidin-biotin complex 면역조직염색을 실시한 결과 CD3에서 강한 양성 반응을 나타냈고 CD4와 CD8에서는 약한 반응을 나타냈다. 조직부위를 보면 피막, 혈관, 임파관, 백수부위와 림프구 및 대식 세포의 세포막에서 양성반응을 보였다.

• 한국수산과학회지
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• 제33권1호
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• pp.55-59
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• 2000

무지개 송어의 뇌하수체 및 배양액에 존재하는 GTH II 농도를 측정하기 위해 Avidin- Biotin complex를 이용한 sandwich EIA 계을 개발했다. Protein A sepharose affinity chromatography을 통해서 얻어진 연어 GTH II의 rabbit IgG에 biotinylation시킨 것 (Biotin-salmon GTH II rabbit IgG)을 제2 항체로 사용하였고, Non-Biotin salmon GTH II rabbit IgG는 단지 protein A sepharose affinity chromatography에서 얻어진 IgG를 제 1 항체로 사용하였다. EIA는 sandwich법에 의해서 이루어졌으며, 효소반응 기질로는 TMB(3,3'5,5-tetramethylbenzidine)를 이용했으며, 반응후 450 nm의 흡광도에서 automatic microplate reader로 측정하였다. 그 결과, $0.12\;{\~}\;125\;ng/ml$의 범위에서 용량반응곡선을 얻었으며, 측정감도 (최소 검출량)는 거의 0.58 ng/ml 정도 였다. 그리고 뇌하수체 추출물 및 배양액 각각의 희석곡선은 GTH II 표존곡선과 일치 하였다. 또한 이러한 GTH II의 표준곡선는 뇌하수체내 다른 peptide hormone와는 교차반응을 거의 나타내지 않았다. Testosterone을 처리한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여 sGnRH에 의한 GTH II 분비량을 본 sandwich EIA계와 RIA계를 비교 조사한 결과, 거의 같은 분비량을 나타냈을 뿐만아니라 같은 분비 pattern을 나타냈다. 이러한 결과로부터 본 sandwich법 EIA계에 의해서 연어과 어류의 뇌하수체 추출물 및 뇌하수체 배양액 중의 GTH II 함량 및 분비량을 측정하는데 있어서 안정된 assay계라고 생각되어진다.

• 대한수의학회지
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• 제38권4호
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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.