• Proceedings of the Korean Nuclear Society Conference
• /
• 2005.05a
• /
• pp.1016-1017
• /
• 2005

• Molecules and Cells
• /
• v.24 no.2
• /
• pp.268-275
• /
• 2007

Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

• Proceedings of the PSK Conference
• /
• 2003.10b
• /
• pp.99.3-100
• /
• 2003

It is desirable to improve the tumor targeting and blood clearance pharmacokinetics of radiolabeled monoclonal antibodies. To achieve this goal, several avidin-biotin (Bt) binding systems have been developed to decouple large molecular weight antibodies from small radiolabels, thereby achieving high tumor-to-background radioactivity ratios. We inserted a readily catabolizable linker, triglycine (TG), between 3-［/sup125/I］iodobenzoate and dendrimer (G3). We also neutralized the positive charges of G3 by acylation with tetrafluorophenyl glycolate, thereby blocking proximal tubular reabsorption of G3 mediated by charge attraction. (omitted)

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.33 no.1
• /
• pp.55-59
• /
• 2000

A specific and sensitive sandwich enzyme-immunoassay (EIA) using Avidin-Biotin complex was developed for the measurement of GTH II levels in pituitary content and pituitary cell culture medium of the rainbow trout-(Oncorhpchus mykiss). Biotin-salmon GTH II rabbit IgG (sefondary antibody) wai purified by a protein A sepharose affinity chromatography column and that was biotinylated by using Biotin-N-hydroxysuccinimide ofter (BNHS). Non-biotin salmon GTH II rabbit IgG (first antibody) was obtained only through a protein A sepharose affinity chromatography column. The assay was performed by the so-called 'sandwich' method using a microtiter plate, A dose-response curve was obtained between $0.12 to 125 ng/ml$ of salmon GTH II. The displacement curves for pituitary extraction and pituitary cell culture medium of testosterone-treated rainbow trout were Parallel to the standard curie. The intra-assay and inter-assay coefficients of variation (CV) were $8.2{\%} (N=5) and 12.5{\%} (N=6)$, respectively, This assay system was used to measure the amount of GTH II that accumulated in the culture medium of dispersed pituitary cells in testosterone-treated immature rainbow trout, The accumulation was increased with the amount or salmon gonadotropin-releasing hormone. GTH II values determined by the present method were well correlated with those determined by radioimmunoassay. As a result, this assay system was found to be suitable for the measurement of GTH II for pituitary extraction and pituitary culture medium in many salmonid fishes.

• Parasites, Hosts and Diseases
• /
• v.30 no.1
• /
• pp.25-32
• /
• 1992

Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as pri-mary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody(MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.

• Tuberculosis and Respiratory Diseases
• /
• v.62 no.1
• /
• pp.43-50
• /
• 2007

Background: Mutated or deregulated expression of C-erbB-2 causes this gene to function as a potent oncogene. Vascular endothelial growth factor (VEGF) is a crucial angiogenic molecule in lung cancer. Both C-erbB-2 and VEGF can promote growth, proliferation and metastasis in non-small cell lung cancer (NSCLC). The purpose of this study was to investigate evaluate the relationship between the expressions of the C-erbB-2 and VEGF genes using immunohistochemistry. Materials and Methods: Ninety-five patients with NSCLC were involved (60 squamous cell carcinoma and 35 adenocarcinoma). The formalin-fixed paraffin embedded specimens were immunohistochemically stained for C-erbB-2 and VEGF using the avidin-biotin complex method. Results: Positive C-erbB-2 expression was observed more often in adenocarcinomas than squamous cell carcinomas (p<0.05). Although the immunohistochemical expressions of C-erbB-2 and VEGF in non-small-cell lung cancer showed increased tendencies at an advanced stage, the correlation between early and advanced cancers was insignificant. In adenocarcinomas, the expressions of VEGF and C-erbB-2 were significantly (p<0.05). Conclusion: The overexpression fo C-erbB-2 was significantly higher in adenocarcinomas than squamous cell carcinomas, and correlated with the expression of VEGF in adenocarcinomas of the lungs.

• Parasites, Hosts and Diseases
• /
• v.29 no.4
• /
• pp.397-402
• /
• 1991

Paragonimus westermani is a common fluke in Korea. The present study aimed to determine serum total IgE and specific IgG levels in experimental paragonimiasis of rats. Each Wistar rat was inoculated orally with 20~25 metacercariae of P. westermani from Cambaroides similis. Before and after infection(1,2,3,4,6,8 weeks) of P. westermani, the blood was collected from the retro- orbital venous plexus of rats and kept serum at $-70^{\circ}C$. Serum total IgE and specific IgG levels were determined by the capture and conventional enzyme-linked immunosorbent assay, respectively. The results were as follows; 1. Serum IgE values were increased to 0. 18${\pm}$0.042 at 2 weeks, $0.28{\pm}0.151$ at 4 weeks and 0.43${\pm}0.055$ at 8 weeks after infection. The absorbances of non-infected rats ranged $0.07{\pm}0.021~0.12{\pm}0.025$. 2. Specific IgG values were slightly increased at 3 weeks ($0.20{\pm}$0.032) and gradually increased up to 8 weeks($0.31{\pm}0.067$) after infection. The absorbances of non-infected rats ranged $0.11{\pm}0.035~0.18{\pm}0.019$. The present results suggested that p. westermani could elevate serum IgE and specific IgG antibodies in Wistar rats which were not a good definitive host.

• Food Science and Biotechnology
• /
• v.16 no.3
• /
• pp.337-342
• /
• 2007

Conventional methods for pathogen detection and identification are labor-intensive and take days to complete. Biosensors have shown great potential for the rapid detection of foodborne pathogens. Fiber-optic biosensors have been used to rapidly detect pathogens because they can be very sensitive and are simple to operate. However, many fiber-optic biosensors rely on manual sensor handling and the sandwich assay, which require more effort and are less sensitive. To increase the simplicity of operation and detection sensitivity, a binding inhibition assay method for detecting Listeria monocytogenes in food samples was developed using an automated, fiber-optic-based immunosensor: RAPTOR (Research International, Monroe, WA, USA). For the assay, fiber-optic biosensors were developed by the immobilization of Listeria antibodies on polystyrene fiber waveguides through a biotin-avidin reaction. Developed fiber-optic biosensors were incorporated into the RAPTOR to evaluate the detection of L. monocytogenes in frankfurter samples. The binding inhibition method combined with RAPTOR was sensitive enough to detect L. monocytogenes ($5.4{\times}10^7\;CFU/mL$) in a frankfurter sample.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.4
• /
• pp.1154-1158
• /
• 2004

The classical ABC (avidin-biotin peroxidase complex) method for immunohistochemistry in the paraffin-embedded tissues bring into being disadvantage such as low sensitivity of antigen detection and highly background. The biotinyl-tyramide conjugation recently introduced for sensitive immunohistochemistry was applied to light microscopy in paraffin-embedded pancreatic and liver tissues. The protocol consists of an indirect method in which 4-5㎛ tissue sections are reacted successively within a specific primary antibody, followed by a biotinylated secondary antibody, streptavidin-horseradich peroxidase (HRP), and then finally with biotinyl-tyramide. The labeling obtained for insulin and collagen antigen tested in pancreatic and liver tissues, respectively, was found to be highly specific with the labeling for each antigen confined to its particular cellular compartment. In this study, fish (flounder) serum was specially applied to remove nonspecific binding. Background levels and nospecific deposition of the staining were negligible. This results suggest that super-signal induction method by biotinyl-tyramide conjugate can readily applied to antigen detection of the paraffin-embedded tissues.

• Korean Journal of Veterinary Research
• /
• v.32 no.2
• /
• pp.227-233
• /
• 1992

Out of twenty rabbits which died of rabbit hemorrhagic disease spontaneously occurring in Korea, five animals had a concurrent infection with Encephalitozoon cuniculi in the cerebrum. The lesions were composed of granulomas, leptomeningitis and perivascular cuffing with mononuclear cells. The granulomas consisted of a central necrotic focus surrounded by an infiltration with plasma cells, lymphocytes and macrophages. Gliosis was associated with the granulomas. Gram-positive organisms were detected in the cerebrum from two rabbits. They were oval to rod-shaped with blunt round ends. The distribution of the pathogens was investigated by the direct avidin-biotin peroxidase complex method. They were present in pseudocysts and macrophages. Pseudocysts were found in the granulomas as well as the neuropil without cellular reactions. Some organisms were present within reticulo-endothelial cells of blood capillaries and macrophages in the subarachnoid spaces. These organisms had ultrastructural characteristics consistent with Encephalitozoon cuniculi.