• KSBB Journal
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• 제22권6호
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• pp.426-432
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• 2007

QDs을 기반으로 하는 OsFIA는 매우 빠르고 간단히 수행될 수 있다. 또한 이 분석법은 고체상의 담체나 결합/잔류시약의 분리 등과 같은 여러 과정을 필요로 하지 않으며, 적은 양의 시약으로도 분석이 가능하다. 본 분석법은 높은 감도로 항원을 측정할 수 있으며, 일상적인 분석에도 쉽게 도입될 수 있을 것이다. 선형 범위 내에서 측정 가능한 receptor의 최소농도는 0.05 nM (2.65 ng/mL) 정도이다. 또한, 일반적으로 상용화된 항체를 가치고 수행이 가능하다. 이 OsFIA 분석법은 기존의 실험적 sandwich immunoassay의 효과적인 대안으로 제시된다.

• Korean Chemical Engineering Research
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• 제59권3호
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• pp.366-372
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• 2021

캐드헤린-카테닌 복합체는 세포의 부착 접합부에서 힘의 전달에 중요한 역할을 하는 것으로 생각된다. 그러나 기계적 힘 신호를 시각화 하고 감지하는 적절한 도구의 부재로, 캐드헤린-카테닌 복합체가 세포 간 접합에서 힘 전달을 조절하는 기본 메커니즘은 아직 파악하기가 어렵다. 본 연구에서는 형광공명에너지전이를 기반으로 설계된 알파카테닌 센서를 사용하여 캐드헤린에 의해 매개되는 힘 전달을 시각화 하였다. 이러한 결과는 알파카테닌이 세포-세포 접합부에서 캐드헤린 매개 기계적에너지변환(mechanotransduction) 경로의 핵심적인 힘 트랜스듀서(force transducer) 임을 보여준다. 본 연구는 향후 기계적 힘의 세포-세포 상호간의 의사소통에 미치는 영향과 생리학적/병리학적 현상과의 관계를 연구하는 데 중요한 이해를 제공할 것이라 본다.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.105-106
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• 2013

Surface plasmon resonance (SPR) is classified into the propagating surface plasmon (PSP) excited on flat metal surfaces and the local surface plasmon (LSP) excited by metalnanoparticles. It is known that fluorescence signals are enhanced by these two SPR-fields.On the other hand, fluorescence is quenched by the energy transfer to metal (FRET). Bothphenomena are controlled by the distance between dyes and metals, and the degree offluorescence enhancement is determined by the correlation. In this study, we determined thecondition to achieve the maximum fluorescence enhancement by adjusting the distance of ametal nanoparticle 2D sheet and a quantum dots 2D sheet by the use of $SiO_2$ spacer layers. The 2D sheets consisting of myristate-capped Ag nanoparticles (AgMy nanosheets) wereprepared at the air-water interface and transferred onto hydrophobized gold thin films basedon the Langmuir-Schaefer (LS) method [1]. The $SiO_2$ sputtered films with different thickness (0~100 nm) were deposited on the AgMy nanosheet as an insulator. TOPO-cappedCdSe/CdZnS/ZnS quantum dots (QDs, ${\lambda}Ex=638nm$) [2] were also transferred onto the $SiO_2$ films by the LS method. The layered structure is schematically shown in Fig. 1. The result of fluorescence measurement is shown in Fig. 2. Without the $SiO_2$ layer, the fluorescence intensity of the layered QD film was lower than that of the original QDs layer, i.e., the quenching by FRET was predominant. When the $SiO_2$ thickness was increased, the fluorescence intensity of the layered QD film was higher than that of the original QDs layer, i.e., the SPR enhancement was predominant. The fluorescence intensity was maximal at the $SiO_2$ thickness of 20 nm, particularly when the LSPR absorption wavelength (${\lambda}=480nm$) was utilized for the excitation. This plasmonic nanosheet can be integrated intogreen or bio-devices as the creation point ofenhanced LSPR field.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.703-706
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• 2000

We constructed a biospecific affinity surface using hyper-branched dendrimers on gold for biospecific recognition, and characterized the resulting surfaces by using confocal fluorescence microscopy. The dendrimer monolayer was firstly constructed on the mercaptoundecanoic acid SAM/Au with pentafluorophenyl ester activation and further functionalized with sulfo-NHS-biotin, an activated ester of biotin. To confirm the formation of biospecific affinity surface, FITC(fluorescein isothiocyanate)-labeled avidin was loaded onto the biotinylated dendrimer monolayer, and fluorescence images of the bound avidins were investigated with a confocal microscope. The constructed biospecific affinity surface showed a much more dense and uniform fluorescence compared to those from poly-L-lysine- and cystamine SAM-based affinity surfaces. For the dependency on the concentration of added FITC-labeled avidin on the affinity surface, derived fluorescence could be detectable from as low as $1{\mu}g/ml$, and intensified up to $50{\mu}g/ml$. Further reaction of FITC-labeled avidin layer with TMR(tetramethylrhodamine)-biocytins resulted in the efficient FRET(fluorescence resonance energy transfer) phenomenon. As an extension of the study, we attempted a patterning of the affinity surfaces on gold by microcontact printing. Fluorescence of the patterned surface demonstrated that FITC-labeled avidin molecules were specifically bound to the biotinylated patches.

• Bulletin of the Korean Chemical Society
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• 제35권7호
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• pp.2114-2122
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• 2014

The association of silybin with ${\beta}$-cyclodextrin and its influence on silybin's binding with bovine serum albumin are reported. The stoichiometry, binding constant, and the structure of silybin-${\beta}$-cyclodextrin inclusion complex are reported. The titrations of silybin with bovine serum albumin in the absence and presence of ${\beta}$-cyclodextrin are carried out and the differences in binding strengths are discussed. Molecular modeling is used to optimize the sites and mode of binding of silybin with bovine serum albumin. F$\ddot{o}$rster resonance energy transfer is calculated and the proximity of interacting molecules is reported in the presence and absence of ${\beta}$-cyclodextrin.

• The Plant Pathology Journal
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• 제34권2호
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• pp.85-92
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• 2018

Enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR), widely used for the detection of plant viruses, are not easily performed, resulting in a demand for an innovative and more efficient diagnostic method. This paper summarizes the characteristics and research trends of biosensors focusing on the physicochemical properties of both interface elements and bioconjugates. In particular, the topological and photophysical properties of quantum dots (QDs) are discussed, along with QD-based biosensors and their practical applications. The QD-based Fluorescence Resonance Energy Transfer (FRET) genosensor, most widely used in the biomolecule detection fields, and QD-based nanosensor for Rev-RRE interaction assay are presented as examples. In recent years, QD-based biosensors have emerged as a new class of sensor and are expected to open opportunities in plant virus detection, but as yet there have been very few practical applications (Table 3). In this article, the details of those cases and their significance for the future of plant virus detection will be discussed.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제45회 하계 정기학술대회 초록집
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• pp.281.2-281.2
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• 2013

Since heavy metal ions included in water or food resources have critical effects on human health, highly sensitive, rapid and selective analysis for heavy metal detection has been extensively explored by means of electrochemical, optical and colorimetric methods. For example, quantum dots (QDs), such as semiconductor QDs, have received enormous attention due to extraordinary optical properties including high fluorescence intensity and its narrow emission peaks, and have been utilized for heavy metal ion detection. However, the semiconductor QDs have a drawback of serious toxicity derived from cadmium, lead and other lethal elements, thereby limiting its application in the environmental screening system. On the other hand, Graphene oxide (GO) has proven its superlative properties of biocompatibility, unique photoluminescence (PL), good quenching efficiency and facile surface modification. Recently, the size of GO was controlled to a few nanometers, enhancing its optical properties to be applied for biological or chemical sensors. Interestingly, the presence of various oxygenous functional groups of GO contributes to opening the band gap of graphene, resulting in a unique PL emission pattern, and the control of the sp2 domain in the sp3 matrix of GO can tune the PL intensity as well as the PL emission wavelength. Herein, we reported a photoluminescent GO array on which heavy metal ion-specific DNA aptamers were immobilized, and sensitive and multiplex heavy metal ion detection was performed utilizing fluorescence resonance energy transfer (FRET) between the photoluminescent monolayered GO and the captured metal ion.

• 생명과학회지
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• 제32권2호
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• pp.148-154
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• 2022

국소접착인산화효소(FAK)는 국소접착부에서 세포부착, 세포이동, 세포역학적 신호전달 등에 관여한다고 알려져 있다. 그러나 세포 외 기질(ECM)과 상호작용하는 인테그린 막단백질과 함께 위치하는 세포막 미세영역(membrane microdomain)의 종류와 ECM 구성에 따른 FAK 활성은 여전히 불분명하다. 형광 공명 에너지 전달(FRET)을 기반으로 유전적으로 인코딩 된 바이오센서는 세포 내 FAK 신호를 높은 시공간 해상도로 제공할 수 있다. 본 연구에서는 유리, 제1형 콜라겐, 피브로넥틴, 라미닌의 ECM 조건에서 FRET 기반 막 표적 FAK 바이오센서를 사용하여 지질유동섬(Lipid raft) 및 비-지질유동섬(non-Lipid raft)에서 FAK의 활성을 분석하고 시각화 하였다. 흥미롭게도, 지질유동섬에서 라미닌 조건 하의 FAK 활성은 다른 ECM 조건보다 낮았고, 비-지질유동섬에서 FAK 활성은 다른 ECM 조건보다 낮았다. 동일한 ECM 조건 상의 비교에서는 피브로넥틴 조건일 때 지질유동섬에서 비-지질유동섬 보다 높은 FAK 활성이 관측되었다. 따라서 이번 연구는 FAK 활성도가 ECM 유형 및 세포막 미세영역에 따라 특이적으로 조절되는 것을 시각적, 정량적으로 보여준다.

• Journal of Crop Science and Biotechnology
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• 제11권2호
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• pp.91-100
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• 2008

Melting curve analysis of fluorescently labeled DNA fragments is used extensively for genotyping single nucleotide polymorphism(SNP). Here, we evaluated a SNP genotyping method by melting curve analysis with the two probe chemistries in a 384-well plate format on a Roche LightCycler 480. The HybProbe chemistry is based on the fluorescence resonance energy transfer(FRET) and the SimpleProbe chemistry uses a terminal self-quenching fluorophore. We evaluated FRET HybProbes and SimpleProbes for two SNP sites closely linked to two quantitative trait loci(QTL) for southern root-knot nematode resistance. These probes were used to genotype the two parents and 94 $F_2$ plants from the cross of PI 96354$\times$Bossier. The SNP genotypes of all samples determined by the LightCycler software agreed with previously determined SSR genotypes and the SNP genotypes determined on a Luminex 100 flow cytometry instrument. Multiplexed HybProbes for the two SNPs showed a 98.4% success rate and 100% concordance between repeats two of the same 96 DNA samples. Also, we developed a HybProbe assay for the Rcs3 gene conditioning broad resistance to the frogeye leaf spot(FLS) disease. The LightCycler 480 provides rapid PCR on 384-well plate and allows simultaneous amplification and analysis in approximately 2 hours without any additional steps after amplification. This allowed for a reduction of the potential contamination of PCR products, simplicity, and enablement of a streamlined workflow. The melting curve analysis on the LightCycler 480 provided high-throughput and rapid SNP genotyping and appears highly effective for marker-assisted selection in soybean.

• Journal of Ginseng Research
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• 제48권1호
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• pp.89-97
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• 2024

Background: Ginsenoside F2 (GF2), the protopanaxadiol-type constituent in Panax ginseng, has been reported to attenuate metabolic dysfunction-associated steatotic liver disease (MASLD). However, the mechanism of action is not fully understood. Here, this study investigates the molecular mechanism by which GF2 regulates MASLD progression through liver X receptor (LXR). Methods: To demonstrate the effect of GF2 on LXR activity, computational modeling of protein-ligand binding, Time-resolved fluorescence resonance energy transfer (TR-FRET) assay for LXR cofactor recruitment, and luciferase reporter assay were performed. LXR agonist T0901317 was used for LXR activation in hepatocytes and macrophages. MASLD was induced by high-fat diet (HFD) feeding with or without GF2 administration in WT and LXRα^-/- mice. Results: Computational modeling showed that GF2 had a high affinity with LXRα. LXRE-luciferase reporter assay with amino acid substitution at the predicted ligand binding site revealed that the S264 residue of LXRα was the crucial interaction site of GF2. TR-FRET assay demonstrated that GF2 suppressed LXRα activity by favoring the binding of corepressors to LXRα while inhibiting the accessibility of coactivators. In vitro, GF2 treatments reduced T0901317-induced fat accumulation and pro-inflammatory cytokine expression in hepatocytes and macrophages, respectively. Consistently, GF2 administration ameliorated hepatic steatohepatitis and improved glucose or insulin tolerance in WT but not in LXRα^-/- mice. Conclusion: GF2 alters the binding affinities of LXRα coregulators, thereby interrupting hepatic steatosis and inflammation in macrophages. Therefore, we propose that GF2 might be a potential therapeutic agent for the intervention in patients with MASLD.