• Title/Summary/Keyword: Label-free detection

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Emission Detection of Mercuric Ions in Aqueous Media Based-on Dehybridization of DNA Duplexes

  • Oh, Byul-Nim;Wu, Qiong;Cha, Mi-Sun;Kang, Hee-Kyung;Kim, Jin-Ah;Kim, Ka-Young;Rajkumar, Eswaran;Kim, Jin-Heung
    • Bulletin of the Korean Chemical Society
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    • v.32 no.9
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    • pp.3223-3228
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    • 2011
  • To quantify the presence of mercuric ions in aqueous solution, double-stranded DNA (dsDNA) of poly(dT) was employed using a light switch compound, $Ru(phen)_2(dppz)^{2+}$ (1) which is reported to intercalate into dsDNA of a right-handed B-form. Addition of mercuric ions induced the dehybridization of poly(dT)${\cdot}$poly(dA) duplexes to form a hairpin structure of poly(dT) at room temperature and the metal-to-ligand charge transfer emission derived from the intercalation of 1 was reduced due to the dehybridization of dsDNA. As the concentration of $Hg^{2+}$ was increased, the emission of 1 progressively decreased. This label-free emission method had a detection limit of 0.2 nM. Other metal ions, such as $K^+$, $Ag^+$, $Ca^{2+}$, $Mg^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Co^{2+}$, $Ni^{2+}$, $Cu^{2+}$, $Cd^{2+}$, $Cr^{3+}$, $Fe^{3+}$, had no significant effect on reducing emission. This emission method can differentiate matched and mismatched poly(dT) sequences based on the emission intensity of dsDNA.

Reduced graphene oxide field-effect transistor for biomolecule detection and study of sensing mechanism

  • Kim, D.J.;Sohn, I.Y.;Kim, D.I.;Yoon, O.J.;Yang, C.W.;Lee, N.E.;Park, J.S.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.431-431
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    • 2011
  • Graphene, two dimensional sheet of sp2-hybridized carbon, has attracted an enormous amount of interest due to excellent electrical, chemical and mechanical properties for the application of transparent conducting films, clean energy devices, field-effect transistors, optoelectronic devices and chemical sensors. Especially, graphene is promising candidate to detect the gas molecules and biomolecules due to the large specific surface area and signal-to-noise ratios. Despite of importance to the disease diagnosis, there are a few reports to demonstrate the graphene- and rGO-FET for biological sensors and the sensing mechanism are not fully understood. Here we describe scalable and facile fabrication of rGO-FET with the capability of label-free, ultrasensitive electrical detection of a cancer biomarker, prostate specific antigen/${\alpha}1$-antichymotrypsin (PSA-ACT) complex, in which the ultrathin rGO sensing channel was simply formed by a uniform self-assembly of two-dimensional rGO nanosheets on aminated pattern generated by inkjet printing. Sensing characteristics of rGO-FET immunosensor showed the highly precise, reliable, and linear shift in the Dirac point with the analyte concentration of PSA-ACT complex and extremely low detection limit as low as 1 fg/ml. We further analyzed the charge doping mechanism, which is the change in the charge carrier in the rGO channel varying by the concentration of biomolecules. Amenability of solution-based scalable fabrication and extremely high performance may enable rGO-FET device as a versatile multiplexed diagnostic biosensor for disease biomarkers.

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Graphene Coated Optical Fiber SPR Biosensor

  • Kim, Jang Ah;Hwang, Taehyun;Dugasani, Sreekantha Reddy;Kulkarni, Atul;Park, Sung Ha;Kim, Taesung
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.401-401
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    • 2014
  • In this study, graphene, the most attractive material today, has been applied to the wavelength-modulated surface plasmon resonance (SPR) sensor. The optical fiber sensor technology is the most fascinating topic because of its several benefits. In addition to this, the SPR phenomenon enables the detection of biomaterials to be label-free, highly sensitive, and accurate. Therefore, the optical fiber SPR sensor has powerful advantages to detect biomaterials. Meanwhile, Graphene shows superior mechanical, electrical, and optical characteristics, so that it has tremendous potential to be applied to any applications. Especially, grapheme has tighter confinement plasmon and relatively long propagation distances, so that it can enhance the light-matter interactions (F. H. L. Koppens, et al., Nano Lett., 2011). Accordingly, we coated graphene on the optical fiber probe which we fabricated to compose the wavelength-modulated SPR sensor (Figure 1.). The graphene film was synthesized via thermal chemical vapor deposition (CVD) process. Synthesized graphene was transferred on the core exposed region of fiber optic by lift-off method. Detected analytes were biotinylated double cross-over DNA structure (DXB) and Streptavidin (SA) as the ligand-receptor binding model. The preliminary results showed the SPR signal shifts for the DXB and SA binding rather than the concentration change.

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Platform Technologies for Research on the G Protein Coupled Receptor: Applications to Drug Discovery Research

  • Lee, Sung-Hou
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.1-8
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    • 2011
  • G-protein coupled receptors (GPCRs) constitute an important class of drug targets and are involved in every aspect of human physiology including sleep regulation, blood pressure, mood, food intake, perception of pain, control of cancer growth, and immune response. Radiometric assays have been the classic method used during the search for potential therapeutics acting at various GPCRs for most GPCR-based drug discovery research programs. An increasing number of diverse small molecules, together with novel GPCR targets identified from genomics efforts, necessitates the use of high-throughput assays with a good sensitivity and specificity. Currently, a wide array of high-throughput tools for research on GPCRs is available and can be used to study receptor-ligand interaction, receptor driven functional response, receptor-receptor interaction,and receptor internalization. Many of the assay technologies are based on luminescence or fluorescence and can be easily applied in cell based models to reduce gaps between in vitro and in vivo studies for drug discovery processes. Especially, cell based models for GPCR can be efficiently employed to deconvolute the integrated information concerning the ligand-receptor-function axis obtained from label-free detection technology. This review covers various platform technologies used for the research of GPCRs, concentrating on the principal, non-radiometric homogeneous assay technologies. As current technology is rapidly advancing, the combination of probe chemistry, optical instruments, and GPCR biology will provide us with many new technologies to apply in the future.

Development of serodiagnostic surface plasmon resonance imaging assay for the detection of antibodies to porcine circovirus type 2

  • Park, Chul;Kim, Bum-Seok;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.1-4
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    • 2011
  • A surface plasmon resonance imaging (SPRI) assay was developed for measuring porcine circovirus type 2 (PCV2) antibody using a recombinant capsid protein as an antigen. The diagnostic potential of SPRI for detecting antibodies to the PCV2 capsid protein was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 70 pig serum samples taken from 6 pig farms. There was a strong positive correlation between the SPRI and ELISA (n = 70, r = 0.911, P<0.01). Therefore, this recombinant capsid protein can be used as an antigen for serological studies, and the SPRI, a label-free and high-throughput method, is expected to be a valuable tool in the serodiagnosis of PCV2 infection.

Fabrication and characteristics of MOSFET protein sensor using gold-black gate (Gold-Black 게이트를 이용한 MOSFET형 단백질 센서의 제조 및 특성)

  • Kim, Min-Suk;Park, Keun-Yong;Kim, Ki-Soo;Kim, Hong-Seok;Bae, Young-Seuk;Choi, Sie-Young
    • Journal of Sensor Science and Technology
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    • v.14 no.3
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    • pp.137-143
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    • 2005
  • Research in the field of biosensor has enormously increased over the recent years. The metal-oxide semiconductor field effect transistor (MOSFET) type protein sensor offers a lot of potential advantages such as small size and weight, the possibility of automatic packaging at wafer level, on-chip integration of biosensor arrays, and the label-free molecular detection. We fabricated MOSFET protein sensor and proposed the gold-black electrode as the gate metal to improve the response. The experimental results showed that the output voltage of MOSFET protein sensor was varied by concentration of albumin proteins and the gold-black gate increased the response up to maximum 13 % because it has the larger surface area than that of planar-gold gate. It means that the expanded gate allows a larger number of ligands on same area, and makes the more albumin proteins adsorbed on gate receptor.

A High-Lateral Resolution MALDI Microprobe Imaging Mass Spectrometer Utilizing an Aspherical Singlet Lens

  • Han, Sang Yun;Kim, Hwan Jin;Ha, Tae Kyung
    • Bulletin of the Korean Chemical Society
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    • v.34 no.1
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    • pp.207-210
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    • 2013
  • We report the construction of a MALDI imaging mass spectrometer equipped with a specially designed laser focusing lens, a compact aspherical singlet lens, that obtains a high-lateral imaging resolution in the microprobe mode. The lens is specially designed to focus the ionization laser (${\lambda}$ = 355 nm) down to a $1{\mu}m$ diameter with a long working distance of 34.5 mm. With the lens being perpendicular to the sample surface and sharing the optical axis with the ion path, the imaging mass spectrometer achieved an imaging resolution of as good as $5{\mu}m$ along with a high detection sensitivity of 100 fmol for peptides. The mass resolution was about 900 (m/${\Delta}m$) in the linear TOF mode. The high-resolution capability of this instrument will provide a new research opportunity for label-free imaging studies of various samples including tissues and biochips, even for the study at a single cell level in the future.

Study on the chemical environment for conformational change of i-motif DNA by atomic force microscopy cantilever (AFM 캔틸레버를 이용한 i-motif DNA의 구조 변화에 미치는 화학적 환경에 대한 연구)

  • Jung, Hwi-Hun;Park, Jin-Sung;Yang, Jae-Moon;Lee, Sang-Woo;Eom, Kil-Ho;Kwon, Tae-Yun;Yoon, Dae-Sung
    • Journal of Sensor Science and Technology
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    • v.19 no.3
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    • pp.214-220
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    • 2010
  • Three-dimensional(3D) structure of specific DNA can be changed between two conformations under an external environmental transition such as pH and salt concentration variations. We have experimentally observed the conformational transitions of i-motif DNA using AFM cantilever bioassay. It is shown that pH change of a solvent induces the bending defleciton change of a cantilever functionalized by i-motif DNA. This indicates that cantilever bioassay enables the label-free detection of DNA structural changes upon pH change. It is implied that cantilever bioassay can be a de novo route to quantitatively understand the conformational transitions of biological molecules under environmental changes.

Control of Optical Hysteresis in Block Copolymer Photonic Gels: A Step Towards Wet Photonic Memory Films

  • Kim, Eun-Ju;Gang, Chang-Jun;Jang, Yu-Rim;Senthilkumar, S.;Gang, Yeong-Jong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.279-279
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    • 2010
  • Smart gels have recently associated with photonic crystals to form photonic gels. Since these photonic gels are capable of reversibly converting the volume change of gels induced in response to external chemical or electric stimuli into characteristic optical signals, they have been considered not only as a good platform for label-free chemical or biological detection, actuators or optical switches but also as a good model system to investigate gel swelling behaviour. Recently, we reported block copolymer photonic gels self-assembled from polystyrene-b-poly(2-vinyl pyridine) (PS-b-P2VP) block copolymers, and demonstrated that selective swelling of lamellar structure allows extremely large tunability of the photonic stop band from UV region to IR region ($\lambda$ peak=350~1,600 nm). Herein we report block copolymer photonic gels which exhibit strong tunable optical hysteresis and their applications. As nonlinear responses in swelling of hydrogels were often observed, photonic gels exhibit optical hysteresis with change of external pH. We demonstrate such optical hysteresis can be precisely programmed by controlling ion-pairing affinity. We anticipate that photonic gels with carefully tunned optical hysteresis are applicable to optical memory devices.

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Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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