• BMB Reports
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• 제30권5호
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• pp.308-314
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• 1997

We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

• Journal of Life Science
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• 제9권1호
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• pp.58-63
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• 1999

Highly metastatic mouse T-lymphoma CS21 cells can grow in vitro when cocultured with CA12 lymph node stromal cells, but they undergo apoptotic cell death when separated from CA12 stromal cells. It has been found that cysteine and interleukin-7(IL-7) as antiapoptotic soluble factors that produced by CA12 stromal cells. In this study, we report that an ICE family protease is activated in CS21 cells when separated from CA12 stromal cells and cultured alone. Enzyme purification using an avidin affinity column revealed that the involved cysteine protease possessed caspase3-like death protease activity. In addition, when IL-7 was added to CS21 cell culture, the protease activity could not be detected during partial purification of the enzyme. Taken together, these results strongly suggest that the caspase3-like protease activation is suppressed by IL-7 as an antiapoptotic factor that leads to abrogation of apoptosis execution.

• Fisheries and Aquatic Sciences
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• 제3권1호
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• pp.52-63
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• 2000

Non-specific reaction has been a problem in doing, especially, research and diagnosis for infectious agents. Avidin-biotin-peroxidase complex (ABC) techniques has widely been used to amplify a reaction. Photobacterium damse1a subsp. piscicdia (formerly Pasteurella piscicida) exhibited a capacity to bind with streptavidin non-specifically. The band, estimated 26 K Da in Western blotted paper, was blocked with biotin but incompletely. In an attempt to explore an involvement of the non-specific substance in attaching piscine cells, cell attachment test performed using anti- Ph. d. subsp piscicida sera raised mouse and rabbit exhibited slightly blocking effects for Mediterranean (1736) and significantly for Japanese (Sp 92144) isolate. Biotin decreased the attachment ability significantly for Sp92144 but it was not effective to 1736. Both isolates showed greatly enhanced attachment ability with poly-L-lysin. The non-specific binding substance was contained in bacterial extracellular products (ECPs). The substance was able to purified with 2-imminobiotin affinity column, the purified substance appeared to have 4 bands in silver staining, and had a carbohydrate branch. This purified substance showed cytotoxic effects selectively between 5 piscine cell lines. Moreover, it stimulated rainbow trout macrophage in terms of reduction of cytochrome cas well as yeast phagocytosis, significantly.

• 동의생리병리학회지
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• 제17권5호
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• pp.1276-1280
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• 2003

To investigate the effect of acupuncture at Zushanli (ST 36) in this study, gastric endocrine cells (G cell) by avidin-biotinylated complex (ABC) technique and histological examinations (HE; periodic acid schiff, PAS; alcian blue stain) of the stomach were perfomed at 1, 3, 6 weeks in normal rats. In other groups, omeprazole were fed for 1, 3, 6 weeks to compare with acupuncture effect. Acupuncture applied to the ST 36 acupoint and the administration of omeprazole increased G cell significantly at 1, 3, 6 weeks in time dependant manner. Furthermore, acupuncture applied to the other acupoint on GB 34 did not produce significant effect. When the common peronial nerve was dissected, acupuncture of ST 36 acupoint produced change of G cell. These data suggest that acupuncture at ST 36 increased G cell in point specific way and that effect was not related with surrounding nerve.

• Journal of Biosystems Engineering
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• 제34권4호
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• pp.254-259
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• 2009

Conventional methods for pathogen detection and identification are labor-intensive and take several days to complete. Recently developed biosensors have shown potential for the rapid detection of foodborne pathogens. In this study, an impedimetric biosensor was developed for rapid detection of Salmonella typhimurium. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on either avidin-biotin binding or self assembled monolayer (SAM) on the surface of the IME to form an active sensing layer. To evaluate effect of antibody immobilization methods on sensitivity of the sensor, detection limit of the biosensor was analyzed with Salmonella samples innoculated in phosphate buffered saline (PBS) or food extract. The impedimetric biosensor based on SAM immobilization method produced better detection limit. The biosensor could detect 107 CFU/mL of Salmonella in pork meat extract. This method may provide a simple, rapid, and sensitive method to detect foodborne pathogens.

• 대한수의학회지
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• 제35권1호
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• pp.29-37
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• 1995

This study was carried out to investigate the morphological changes of the oxytocin secreting neurons in age-related rat hypothalamus by means of immunohistochemistry. The experimental group was 20 aged female rats (Sprague-Dawley, 25~30 months), and the control group was 10 adult female rats (10 months). All animals were perfused transcardially with 4% paraformaldehyde-lysine-periocdate(PLP), and serial transverse brain sections($30{\mu}m$) were prepared by means of cryotome, and were stained with rabbit anti-oxytocin antisera immunohistochemically, using the free floating method and avidin-viotin peroxidase complex. The results were as followings. 1. The immunostained oxytocin secreting neurons were located at the paraventricular nucleus and supraoptic nucleus of age-related rat hypothalamus chiefly. 2. The numbers of oxytocin secreting neurons decreased at the paraventricular nucleus and supraoptic nucleus of age-related rat hypothalamus(p<0.01) 3. The oxytocin secreting neurons of age-related rat hypothalamus immunostained less than those of the adult rat hypothalamus, and the paraventricular nucleus immunostained greater than supraoptic nucleus in age-related rat. 4. The numbers and expansion of dendrite and axonal varicosities decreased in the age-related rat hypothalamus greatly.

• 대한수의학회지
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• 제32권1호
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• pp.91-98
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• 1992

Listeria monocytogenes antigens were detected with the avidinbiotinperoxidase complex(ABPC) method in formalin-fixed, paraffin-embedded tissues from experimentally infected rats, mice and guinea pigs. The anti-Lirteria monocytogenes serum used as first antibody was prepared by immunizing rabbits with Listeria monocytogenes serotype 1/2a. Rats, mice and guinea pigs that had been given inoculation of L monocytogenes(serotype 4b, Scott A strain) via intraperitoneally allotted to 3 groups. Rats were pretreated with the dexamethasone(DM-rats) for 7 consecutive days, mice and guinea pigs were inoculated intraperitoneally with L. monocytogenes At necropsy white necrotic foci of the liver, spleen and kidney were seen in mice and DM-rats, whereas not in guinea pigs. Organisms stained by the ABPC method were identified as pleomorphic dark brown staining structures in the livers, spleens and kidneys of mice and DM-rats. They were present in high numbers in center and peripherial regions of necrobiotic and necrotic foci of the liver and spleen as well as in glomerulus of the renal cortex. and liable tool for confirmative diagnosis of these bacterial diseases.

• 대한수의학회지
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• 제37권1호
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• pp.9-13
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• 1997

ABC법에 의해 면역조직화학적으로 아프리카발톱두꺼비(Xenopus laevis)의 위장관내분비세포를 관찰하였던 바, neurotensin 면역반응세포는 중등도로 소장에서 약하게 반응하였다. 구형의 gastrin releasing peptide(GRP) 면역반응세포는 위에서 위저부의 상피 세포 바로 아래와 유문부선에서 다수로, 소장에서는 가는 방추형으로 극소수 또는 중등도로 분포하였다. 한편 substance P 면역반응세포는 방추형으로 장상피에서 관찰되었다. 그러나 secretin, motilin, met-enkephalin-8 (M-Enk) 및 polypeptide YY(PYY) 면역반응세포는 전체 소화관에서 관찰할 수 없었다.

• Bulletin of the Korean Chemical Society
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• 제24권1호
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• pp.70-74
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• 2003

The heterogeneous bioluminescence immunoassay for digoxin was developed using photoprotein, native aequorin as a label and the site-directed immobilization technique based on avidin/biotin interaction. Aequorin is a bioluminescence protein, originally isolated from the jellyfish Aequoria Victoria and an attractive label in analytical applications because of sensitive detection due to virtually no background bioluminescent signal. Digoxin is a cardioactive drug, and its therapeutic level in serum is at low concentration with very narrow therapeutic index. The aequorin-digoxigenin conjugates were synthesized by the N-hydroxysuccinimide ester method and characterized in terms of bioluminescent residual activity. The resulting dose-response curve shows that the detection limit is $1.0\;{\times}\;10^{-10}\;M$ and a dynamic range is three orders of magnitude, which was obtained by $1.0\;{times}\;10^{-10}\;M$ conjugate and 0.9 μg/mL anti-digoxin antibody. Three structurally similar molecules to digoxin were examined for their cross-reactivity. None of these three compounds showed any crossreactivity with digoxin antibody employed in this study. Standard amounts of digoxin corresponding to the therapeutic range were spiked into the each serum solution. Study of the serum matrix effect indicated that correlation coefficient shows good agreement between luminescence light intensity between in buffer and in serum.

• 대한기관식도과학회:학술대회논문집
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• 대한기관식도과학회 1993년도 제27차 학술대회 초록집
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• pp.88-88
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• 1993

현재 악성종양의 조직학적 진단에 면역조직화학반응의 검토가 많이 사용되는데 정상상피가 암으로의 진행시 점막상피세포의 주구성 단백성분에 있는 cytokeratin의 발현성에 변화가 일어난다고 알려져 있다. 이 cytokeratin은 현재 19종의 subclass가 알려져 있는데 각 subclass에 대응하는 항체를 이용하여 후두암세포의 기원을 알고 암조직의 분화도를 판정하고자 본 연구를 시행하였다. 연구방법은 정상성대조직 5예, 성대에 국한된 편평세포암 22예를 monoclonal anti-ck Ab와 polyclonal anti-ck Ab를 사용, Avidin-Biotin Peroxidase Complex method로 면역조직화학적 염색을 시행하였다. 그결과 정상후두상피에서 저분자량의 keratin은 표층을 제외한 심층부에서 양성반응을 보였다. 후두의 편평세포암은 상피성종양이며, 저분자량의 항ck 항체는 저분화암에서 양성반응을 보이고 고분자량의 항ck항체는 고분화암에서 양성반응을 보였다.