• Applied Chemistry for Engineering
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• v.27 no.2
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• pp.185-189
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• 2016

Among many kinds of bioaffinity sensors, the avidin-biotin system has been widely used in a variety of biological applications due to the specific and high affinity interaction of the system. In this work, gold nanorods with high surface area were explored as electrodes in order to amplify the signal response from the avidin-biotin interaction which can be further utilized for avidin-biotin biosensors. Electrochemical performance of electrodes modified with nanorods and functionalized with avidin in response to interactions with biotin at various concentrations using $[Fe(CN)_6]^{3-/4-}$ couple as the redox probe were investigated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). A very low biotin concentration of less than 1 ng/mL could be detected using the electrodes modified with nanorods.

• 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.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.31 no.10
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• pp.986-992
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• 2007

This paper presents the fabrication and characterization of carbon films pyrolyzed with various photoresists for bioMEMS applications. To verify the usefulness of pyrolyzed carbon films as an electrode material in an electrochemical biosensor developed by the authors, interactions between avidin and biotin on the pyrolyzed carbon film were studied via electrochemical impedance spectroscopy based on electrostatic interactions between avidin and negatively-charged ferricyanide. The pyrolyzed carbon films were characterized using a surface profiler, a precision semiconductor parameter analyzer, a nanoindentor, scanning electron microscopy, and atomic force microscopy. Amine conjugated biotin was immobilized on the electrode using EDC/NHS as crosslinkers after $O_2$ plasma treatment to enhance functional groups on the carbon electrode pyrolyzed at $1000^{\circ}C$ with AZ9260. The detection of avidin binding with different concentrations in a range of 0.75 nM to $7.5\;{\mu}M$ to the pyrolyzed carbon electrode modified with biotin was carried out by measuring the electrochemical impedance change. The results show that avidin binds to the biotin on the electrode not by non-specific interaction but by specific interaction, and that EIS successfully detects this binding event. Pyrolyzed carbon films are a promising material for miniaturization, integration, and low-cost fabrication in electrochemical biosensors.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.4
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• pp.263-267
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• 2003

Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.

• 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.

• Journal of Integrative Natural Science
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• v.3 no.1
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• pp.33-37
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• 2010

The functionalized photonic crystals of porous silicon biosensor was prepared for the application as a label-free biosensor based on distributed Bragg reflector interferometer. Prepared distributed Bragg reflector of porous silicon biosensor displayed sharp reflection in the optical reflective spectra. The mean of construction of molecular architectures on distributed Bragg reflector of porous silicon surfaces was investigated for the step-by-step binding interaction with amines, biotin, avidin, and biotinylated protein A. The subsequent introduction of avidin, and biotinylated protein A resulted in the reflectivity shifted to longer wavelengths, indicative of a change in refractive indices induced by binding of biomolecules.

• 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.

• 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.

• Bulletin of the Korean Chemical Society
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• v.24 no.1
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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.

• Journal of Chest Surgery
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• v.39 no.11 s.268
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• pp.828-837
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• 2006

Background: Tissue hypoxia is characteristic of many human malignant neoplasm, and hypoxia inducible factor-1(HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increasing $O_2$ delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-$1{\alpha}$ has been reported in many human malignancies, but in non-small cell lung carcinoma the influence of HIF-$1{\alpha}$ on tumor biology, including neovascularization, is not still defined. In present study the relationship of HIF-$1{\alpha}$ expression on angiogenetic factors, relationship between the tumor proliferation and HIF-$1{\alpha}$ expression, interaction of HIF-$1{\alpha}$ expression and p53, and relationship between HIF-$1{\alpha}$ expression and clinico-pathological prognostic parameters were investigated. Material and Method: Archival tissue blocks recruited in this study were retrieved from fifty-nine patients with primary non-small cell lung carcinoma, who underwent pneumonectomy or lobectomy from 1997 to 1999. HIF-$1{\alpha}$, VEGF(vascular endothelial growth factor), and p53 protein expression and Ki-67 labeling index in tumor tissues were evaluated, using a standard avidin-biotin-peroxidase complex(ABC) immunohistochemistry. Relationship between the HIF-$1{\alpha}$ expression and VEGF, p53 overexpression and correlation between the HIF-$1{\alpha}$ expresseion and Ki-67 index were analyzed. Clinico-pathologic prognostic parameters were also analyzed. Result: HIF-$1{\alpha}$ expression in cancer cells was found in 24 of 59 cases of non-small cell lung carcinoma(40.7%). High HIF-$1{\alpha}$ expression was significantly associated with several pathological parameters, such as pathological TMN stage(p=0.004), pT stage(p=0.020), pN stage (p=0.029), and lymphovascular invasion(p=0.019). High HIF-$1{\alpha}$ expression was also significantly associated with VEGF immunoreactivity(p<0.001), and aberrant p53 expression(p=0.040). but was marginally associated with Ki-67 labeling index(p=0.092). The overall 5-year survival rate was 42.3%. The survival curve of patients with a high HIF-$1{\alpha}$ expression was worse than that of patients with low-expression(p=0.002). High HIF-$1{\alpha}$ expression was independent unfavorable factors with a marginal significance in multivariate analysis performed by Cox regression. Conclusion: It is suggested that high HIF-$1{\alpha}$ expression may be associated with intratumoral neovascularization possibly through HIF-VEGF pathway, and high HIF-$1{\alpha}$ expression could be associated with lymph node metastasis and post operative poor prognosis in patients with non-small cell lung carcinoma.