• Korean Journal of Veterinary Research
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• v.37 no.4
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• pp.825-829
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• 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.

• Bulletin of the Korean Chemical Society
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• v.35 no.2
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• pp.383-391
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• 2014

Biotin-S-S-Phosphine was designed and synthesized as a potential tool for a proteomic study of O-GlcNAcmodified proteins. This reagent features a disulfide linker between a triarylphosphine moiety, which allows selective conjugation to azide-containing proteins, and a biotin moiety that can allow easy isolation through its strong affinity toward avidin-coated solid beads. The disulfide linkage within this reagent can allow the easy release of the bound molecules of interest, which is difficult to achieve when a biotin:avidin pair is used alone, by reducing the disulfide bond of the reagent with DTT. Preliminary in vitro biological assays with azidelabeled and unlabeled cell lysates and a pure protein Nup62 showed that the Biotin-S-S-Phosphine reagent is highly reactive toward the free thiol groups of proteins. When a molecular tool with a disulfide linker is applied to the enrichment of the molecules of interest from other species, it is important to block the free-thiols of the sample using exhaustive alkylation prior to the Staudinger ligation reactions to restore the bioorthogonal nature of this reaction.

• ETRI Journal
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• v.31 no.6
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• pp.647-652
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• 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.

• Bulletin of the Korean Chemical Society
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• v.25 no.1
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• pp.115-118
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• 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.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.52-55
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• 2000

Structually organized mono- and multilayers were developed on gold for the catalytic and affinity biosensing using hyper-branched dendrimers. For the catalytic biosensing interface, a new approach to construct a multilayered enzyme film on the electrode surface was developed. The film was prepared by layer-by-layer depositions of dendrimers and periodate-oxidized glucose oxidase. The voltammograms obtained from the GOx/dendrimer multilayered electrodes revealed that bioelectrocatalytic response is directly correlated to the number of deposited bilayers. From the analysis of voltammetric and ellipsometric signals, the coverage of active enzyme per layer during the layering steps was estimated, demonstrating the spatially-ordered multilayer formation. As an extension of the study, dendrimers having various degrees of ferrocenyl modification were prepared and used. The resulting electrodes were electrochemically characterized, and the density of ferrocenyl groups, active enzyme coverage, and sensitivity were estimated. For the affinity-sensing surrface, a biosensor system based on avidin-biotin interaction was developed. As the building block of affinity monolayer, G4 dendrimer having partial ferrocenyl-tethered surface groups was prepared and used. And the biotinylated and electroactive dendritic monolayer was used for the affinity-sensing surface interacting with avidin. Electrochemical characterization of the resulting biosensor was conducted using free enzyme in electrolyte in terms of degree of surface coverage with avidin and subsequent surface shielding.

• Journal of Pharmaceutical Investigation
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• v.29 no.2
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• pp.87-92
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• 1999

Drug delivery to the brain may be achieved by producing chimeric peptide, attaching the drug to protein 'vectors' which are transported into the brain from the blood by a receptor-mediated transcytosis through the blood-brain barrier (BBB). Since the BBB expresses high concentrations of transferrin receptor, and it was reported that anti-transferrin receptor mouse monoclonal antibody (OX26) undergoes transcytosis through the BBB, it is logical to assume that a drug delivery system via transferrin receptor-mediated transcytosis is a promising strategy. In the present study, therefore, we tested feasibility of several OX26 based vectors for the brain delivery of a model drug. Avidin-based delivery vectors such as OX26-streptavidin (OX26-SA), OX26-neutralite avidin (OX26-NLA) were chemically synthesized vectors and OX26 immunoglobulin G 3 type $C_{H}3$ fusion avidin $(OX26\;IgG3C_H3-AV)$ was genetically engineered. To improve the efficiency of producing chimeric peptide, we used avidin-biotin technology. Pharmacokinetics of $[^3H]biotin$ bound to OX26-SA, OX26-NLA and $OX26\;IgG3C_H3-AV$ was determined by intravenous injection technique, and their stabilities in plasma were analyzed using HPLC. The brain delivery of $[^3H]biotin$ bound to OX26-SA, OX26-NLA and OX26\;$IgG3C_{H}3-AV$ (expressed as %ID/g brain) was $0.22{\pm}0.01$, $0.18{\pm}0.01$ and $0.25{\pm}0.09$, respectively. The areas under the plasma concentration versus time curve (AUC) for OX26-SA, OX26-NLA, $OX26\;IgG3C_H3-AV$ from time zero to 60 min were $209{\pm}10$, $195{\pm}9$, $134{\pm}29\;%ID\;min/ml$ respectively and their total clearances $(CL_{tot})$ were $1.00{\pm}0.09$, $1.08{\pm}0.07$ and $1.54{\pm}0.29\;ml/min/kg$, espectively. These results showed that these vectors possess preferable pharmaceutical (e.g., resonable stability) and pharmacokinetics (e.g., significant brain uptake and enhanced AUC) for brain delivery. Therefore, these vectors may be broadly useful in the brain delivery of drugs that are not transported into the brain to a significant extent.

• Analytical Science and Technology
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• v.23 no.6
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• pp.537-543
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• 2010

Dopamine (DA) is an important neurotransmitter molecule of catecholamines. Its deficiency could lead to brain disorder such as Parkinson's disease and schizophrenia. Therefore, it is necessary to establish a suitable analytical technique with sensitivity and simplicity. A competitive enzyme-linked immunosorbent assay for DA has been optimized and characterized. Assay sensitivity is controlled by two factors in competitive immunoassay. One is a nature and concentration of competitor, and the other is those of binder, antibody. Thus, optimization was performed: BSA-DA conjugate and antibody-avidin conjugate were prepared by dual heterobifunctional coupling method using SATA and SMCC. Assay condition was optimized with $6.66\;{\mu}gmL^{-1}$ of BSA-DA and $4.17{\times}10^{-10}\;M$ of antibody-avidin conjugate. A dose-response curve was constructed, and a limit of detection and a dynamic range for DA were accomplished to $2.3{\times}10^{-2}\;{\mu}g\;mL^{-1}$ and four orders of magnitude ($1.0{\times}10^{-7}\;M$ to $1.0{\times}10^{-3}\;M$), respectively. Calibration curve was constructed on dynamic range and least-squares regression of this data gave the following relationship: absorbance = -0.1098 log[DA]+0.0353 ($R^2$ = 0.9956).

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.107-111
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• 2004

A comparative study was performed to evaluate the signal amplification strategies in electrochemical affinity sensing, which included the direct electron transfer and diffusible-group mediated electron transfer between label enzymes that were specifically bound to target proteins and chemically modified electrode surfaces. As a platform surface for affinity recognition reactions, a double functionalized poly(amidoamine) dendrimer monolayer that was modified with ferrocene and biotin groups was constructed on a gold surface. With the chemically modified electrode, a model affinity sensing with avidin was investigated. The advantages of adopting the diffusible-group mediated signaling strategy were demonstrated in terms of signal sensitivity and stability.

• Bulletin of the Korean Chemical Society
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• v.31 no.5
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• pp.1223-1227
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• 2010

In this study, the orientational behavior of thermotropic liquid crystals (LC) supported on a film of protein receptors was examined. Avidin was roller printed and covalently immobilized onto the surface of gold using NHS/EDC chemistry. The orientation of nematic 4-cyano-4'-pentylbiphenyl (5CB) was found to be parallel to the plane of the printed avidin surface before incubation with a solution of biotin. However, protein-receptor complexation induced a random orientation of 5CB, where protein-receptor complexes disturbed the nanoscale topography of the printed protein surface. Atomic force microscopy and ellipsometry was used to confirm printing and the specific interaction of proteins. These results demonstrate that the combination of LC and roller printing can be used to detect specific interactions between biomolecules by manipulating the orientational behavior of LC to the printed protein surfaces.

• Korean Journal of Veterinary Research
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• v.33 no.1
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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.