Diffusion-Enhanced Lanthanide Nanoparticle FRET Assays
扩散增强型镧系元素纳米粒子 FRET 测定
基本信息
- 批准号:9095386
- 负责人:
- 金额:$ 19.19万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2014
- 资助国家:美国
- 起止时间:2014-08-01 至 2018-04-30
- 项目状态:已结题
- 来源:
- 关键词:AffectAmino AcidsAntibodiesAntigensBee VenomsBindingBiologicalBiological AssayBloodCapsid ProteinsChelating AgentsClinicalCollaborationsComplexComputer softwareDataDetectionDiagnosticDiffuseDiffusionDigoxinDrug ReceptorsElectronicsEpitopesExtinction (Psychology)Fab ImmunoglobulinsFluorescenceFluorescence Polarization ImmunoassayFluorescence Resonance Energy TransferFluorescence SpectroscopyGeneric DrugsGoalsGoldHIVHIV AntibodiesHIV Envelope Protein gp120HealthHumanInstitutesIonsLabelLanthanoid Series ElementsLasersLightMeasurementMeasuresMedicalMembraneMetalsMethodsMicroscopeModelingMolecularN-terminalOpticsOxidesParticle SizePeptidesPharmaceutical PreparationsProteinsProteomicsReactionReaderRhodamineSamplingScanningSensitivity and SpecificitySerumSilverSourceSpectrum AnalysisStagingSubstance of AbuseSurfaceSystemTestingTherapeuticTimeVaccinesVirionWorkabsorptionbasebiological researchdrug discoveryfluorophoremacromoleculemillisecondnanomolarnanoparticlenanosecondneutralizing antibodynovelnovel strategiesparticlereceptorresearch clinical testingscreeningsmall moleculevirologyviron
项目摘要
DESCRIPTION (provided by applicant): We propose to develop a new approach to measuring biomolecule association reactions. This method will be based on diffusion-enhanced FRET (DE-FRET). With nanosecond decay time fluorophores diffusion has essentially no effect on the extent of FRET. We will overcome this limitation by using lanthanide oxide nanoparticles (LNP) which have surface-bound silver particles. The presence of multiple lanthanide ions per particle and Ag particles will compensate for the typical low extinction and intensity of lanthanides, and also circumvent the need for sensitizing chelators. We will use the LNPs as donors and acceptor-labeled detection molecules. Binding of the detection molecules to a target will alter the acceptor diffusion coefficient, the LNPs intensity and decay times. The DE-FRET data will reveal translational diffusion coefficients. Similar information can be obtained by FCS, but FCS requires careful control of the focal volume and can only be used at nanomolar concentrations. In contrast our method will require no special optics, will work with any volume sample and will be sensitive to binding constants in the micromolar range, which is ideal for binding to receptors and drug discovery. We will demonstrate the usefulness of our approach using two types of binding reactions. The first will be self-association of a protein melittin to form tetramer. Measurements of protein- protein association reactions are important for proteomics. The second will be a clinical testing assay of digoxin in serum. This application is representative of large class of small molecule assays of drugs and substances of abuse. In this revised proposal we included an important biomedical application, screening of antibodies against HIV pseudo-virons or gp120. This project will have a high impact because the method can be immediately used with some commercial plate readers and easily added to most existing plate readers.
描述(由申请人提供):我们建议开发一种测量生物分子结合反应的新方法。这种方法将基于扩散增强FRET (DE-FRET)。随着纳秒级的衰减时间,荧光团的扩散基本上对FRET的程度没有影响。我们将通过使用表面结合银粒子的氧化镧纳米粒子(LNP)来克服这一限制。每个粒子和银粒子的多个镧系离子的存在将补偿镧系元素典型的低消光和强度,也规避了敏化螯合剂的需要。我们将使用LNPs作为供体和受体标记的检测分子。检测分子与目标的结合将改变受体扩散系数、LNPs强度和衰减时间。DE-FRET数据将揭示平动扩散系数。FCS可以获得类似的信息,但FCS需要仔细控制焦点体积,并且只能在纳摩尔浓度下使用。相比之下,我们的方法不需要特殊的光学器件,可以处理任何体积的样品,并且对微摩尔范围内的结合常数敏感,这对于结合受体和药物发现是理想的。我们将用两种类型的结合反应来证明我们的方法的有效性。首先是一种蛋白蜂毒素的自结合形成四聚体。蛋白质-蛋白质结合反应的测量对蛋白质组学非常重要。第二项将是血清中地高辛的临床检测。该应用是一类具有代表性的小分子分析药物和滥用物质。在这个修改后的提案中,我们包括了一个重要的生物医学应用,HIV伪病毒或gp120抗体的筛选。这个项目将有很大的影响,因为该方法可以立即与一些商业车牌阅读器一起使用,并且很容易添加到大多数现有的车牌阅读器中。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(1)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Joseph R. LAKOWICZ其他文献
Joseph R. LAKOWICZ的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Joseph R. LAKOWICZ', 18)}}的其他基金
Photonics-based Fluorescence Imaging for Research, Diagnostics, and Pathology
用于研究、诊断和病理学的基于光子学的荧光成像
- 批准号:
10546493 - 财政年份:2022
- 资助金额:
$ 19.19万 - 项目类别:
Photonics-based Fluorescence Imaging for Research, Diagnostics, and Pathology
用于研究、诊断和病理学的基于光子学的荧光成像
- 批准号:
10329143 - 财政年份:2022
- 资助金额:
$ 19.19万 - 项目类别:
Coupled Emission Microscopy for the Biosciences
用于生物科学的耦合发射显微镜
- 批准号:
9424262 - 财政年份:2018
- 资助金额:
$ 19.19万 - 项目类别:
Plasmon-coupled Fluorescence Correlation Spectroscopy in Nanoholes
纳米孔中的等离子体激元耦合荧光相关光谱
- 批准号:
9766321 - 财政年份:2018
- 资助金额:
$ 19.19万 - 项目类别:
Coupled Emission Microscopy for the Biosciences
用于生物科学的耦合发射显微镜
- 批准号:
10093077 - 财政年份:2018
- 资助金额:
$ 19.19万 - 项目类别:
Bioaffinity Assays Using UV One-Dimensional Photonic Crystals (1DPC)
使用紫外一维光子晶体 (1DPC) 进行生物亲和力测定
- 批准号:
9098709 - 财政年份:2015
- 资助金额:
$ 19.19万 - 项目类别:
Multi-User Time-Resolved Fluorescence Spectrometer
多用户时间分辨荧光光谱仪
- 批准号:
8825781 - 财政年份:2015
- 资助金额:
$ 19.19万 - 项目类别:
Bioaffinity Assays Using UV One-Dimensional Photonic Crystals (1DPC)
使用紫外一维光子晶体 (1DPC) 进行生物亲和力测定
- 批准号:
8957305 - 财政年份:2015
- 资助金额:
$ 19.19万 - 项目类别:
Fluorescence Lifetime Imaging Microscopy (FLIM)
荧光寿命成像显微镜 (FLIM)
- 批准号:
7791919 - 财政年份:2010
- 资助金额:
$ 19.19万 - 项目类别:
Sub-Wavelength Imaging of Intracellular Metal Ions
细胞内金属离子的亚波长成像
- 批准号:
7940807 - 财政年份:2009
- 资助金额:
$ 19.19万 - 项目类别:
相似海外基金
Double Incorporation of Non-Canonical Amino Acids in an Animal and its Application for Precise and Independent Optical Control of Two Target Genes
动物体内非规范氨基酸的双重掺入及其在两个靶基因精确独立光学控制中的应用
- 批准号:
BB/Y006380/1 - 财政年份:2024
- 资助金额:
$ 19.19万 - 项目类别:
Research Grant
Quantifying L-amino acids in Ryugu to constrain the source of L-amino acids in life on Earth
量化 Ryugu 中的 L-氨基酸以限制地球生命中 L-氨基酸的来源
- 批准号:
24K17112 - 财政年份:2024
- 资助金额:
$ 19.19万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Collaborative Research: RUI: Elucidating Design Rules for non-NRPS Incorporation of Amino Acids on Polyketide Scaffolds
合作研究:RUI:阐明聚酮化合物支架上非 NRPS 氨基酸掺入的设计规则
- 批准号:
2300890 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Continuing Grant
Basic research toward therapeutic strategies for stress-induced chronic pain with non-natural amino acids
非天然氨基酸治疗应激性慢性疼痛策略的基础研究
- 批准号:
23K06918 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Molecular mechanisms how arrestins that modulate localization of glucose transporters are phosphorylated in response to amino acids
调节葡萄糖转运蛋白定位的抑制蛋白如何响应氨基酸而被磷酸化的分子机制
- 批准号:
23K05758 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Molecular recognition and enantioselective reaction of amino acids
氨基酸的分子识别和对映选择性反应
- 批准号:
23K04668 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Design and Synthesis of Fluorescent Amino Acids: Novel Tools for Biological Imaging
荧光氨基酸的设计与合成:生物成像的新工具
- 批准号:
2888395 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Studentship
Structurally engineered N-acyl amino acids for the treatment of NASH
用于治疗 NASH 的结构工程 N-酰基氨基酸
- 批准号:
10761044 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Lifestyle, branched-chain amino acids, and cardiovascular risk factors: a randomized trial
生活方式、支链氨基酸和心血管危险因素:一项随机试验
- 批准号:
10728925 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别:
Single-molecule protein sequencing by barcoding of N-terminal amino acids
通过 N 端氨基酸条形码进行单分子蛋白质测序
- 批准号:
10757309 - 财政年份:2023
- 资助金额:
$ 19.19万 - 项目类别: