PEPTIDE BEAD MICROARRAYS MEASURED BY MASS SPECTROMETRY IMAGING

通过质谱成像测量肽珠微阵列

• 批准号: 8312025
• 负责人: Vladislav Bergo
• 金额: $ 34.9万
• 依托单位: ADEPTRIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-28 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8312025
• 关键词: Adopted; Affinity; Antibodies; Area; Biochemical; Biological; Biological Assay; Biomedical Research; Businesses; Caliber; Chemistry; Clinical; Computer software; Data; Data Quality; Detection; Development; Diagnostic; Ensure; Enzymes; Epitopes; Equipment; Evaluation; Fluorescence; Image; Image Analysis; Individual; Industry; Label; Libraries; Link; MALDI-TOF Mass Spectrometry; Manufacturer Name; Mass Spectrum Analysis; Measurement; Measures; Methods; Microarray Analysis; Microspheres; Modification; Peptide Hydrolases; Peptide Library; Peptides; Performance; Phase; Plant Resins; Preparation; Proteins; Proteomics; Protocols documentation; Reaction; Reagent; Relative (related person); Research; Running; Sampling; Scanning; Screening procedure; Services; Site; Slide; Solid; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spottings; Statistical Data Interpretation; Surface; Suspension substance; Suspensions; Techniques; Technology; Testing; Time; Tissues; base; combinatorial; cost; design; enzyme activity; innovation; meetings; member; novel; research study; tool

DESCRIPTION (provided by applicant): New proteomic technologies are urgently needed that can meet the demand of emerging large-scale biological studies. We have developed a novel microarray analysis platform, which enables screening of large bead libraries using MALDI TOF mass spectrometry imaging. While biological tissue imaging by mass spectrometry is now a well-established technique, MALDI TOF MSI has not been used to perform scanning of biological microarrays. Our MSBeadArray technology enables transfer of selected analytes from beads onto the surface of a specially designed microarray slide under conditions, which maintain spatial separation of analytes from individual microbeads while preserving co-localization of analytes extracted from the same bead. The unique features of this platform include MS analysis of bead libraries containing up to 500,000 samples on a single microarray slide, dual fluorescence - MSI readout and the option to perform label-free sample detection that eliminates the need for fluorescent labels and detection antibodies. Here we propose to apply the MSBeadArray technology to the development of libraries of bead-conjugated peptides, which will be measured by mass spectrometry in a microarray format. Peptide bead arrays with the mass spectrometry readout will have significant advantage over both planar peptide microarrays with positional encoding and suspension bead arrays with the fluorescent encoding (e.g., the Luminex" platform). Because mass spectrometry can detect multiple peptide species in a single measurement, the peptide purity, presence of additional sequences and even relative amounts of each peptide per bead are measured during the microarray scan. Peptide manufacturers routinely employ MALDI TOF MS analysis to perform QC of the synthesized peptides that are used to produce microarrays. However the fabricated microarrays are not analyzed by MS, therefore the quality of peptide probes, immobilized on the microarray slides or beads, is not known. In contrast, our technology performs QC on every member of the bead microarray after the microarray reaction is completed. Variability of bead assays will be substantially reduced because of the probe quality data provided by the MSBeadArray platform. PUBLIC HEALTH RELEVANCE: New proteomic technologies are urgently needed that can meet the demand of emerging large-scale biological studies. We have developed a novel microarray analysis platform, which enables screening of large bead libraries using MALDI TOF mass spectrometry imaging. Here we propose to apply the MSBeadArray technology to the development of libraries of bead-conjugated peptides, which will be measured by mass spectrometry in a microarray format.

描述（由申请人提供）：迫切需要新的蛋白质组学技术，以满足新兴的大规模生物学研究的需求。我们已经开发了一种新的微阵列分析平台，它可以使用MALDI TOF质谱成像筛选大珠库。虽然通过质谱的生物组织成像现在是一种成熟的技术，但MALDI TOF MSI尚未用于进行生物微阵列的扫描。我们的MSBeadArray技术能够在特定条件下将选定的分析物从微珠转移到专门设计的微阵列载玻片表面上，该条件保持分析物与单个微珠的空间分离，同时保持从同一微珠提取的分析物的共定位。该平台的独特功能包括在单个微阵列载玻片上对含有多达500，000个样品的珠库进行MS分析，双重荧光- MSI读数以及执行无标记样品检测的选项，从而消除了对荧光标记和检测抗体的需要。在这里，我们建议将MSBeadArray技术应用于珠缀合肽的文库的开发，所述珠缀合肽将以微阵列形式通过质谱法测量。具有质谱读数的肽珠阵列将具有优于具有位置编码的平面肽微阵列和具有荧光编码的悬浮珠阵列（例如，Luminex”平台）。因为质谱法可以在单次测量中检测多个肽种类，所以在微阵列扫描期间测量肽纯度、额外序列的存在以及甚至每个珠粒的每种肽的相对量。肽生产商通常使用MALDI TOF MS分析来对用于生产微阵列的合成肽进行QC。然而，制造的微阵列不通过MS分析，因此固定在微阵列载玻片或珠上的肽探针的质量是未知的。相比之下，我们的技术在微阵列反应完成后对微珠微阵列的每个成员进行QC。由于MSBeadArray平台提供的探针质量数据，微珠检测的变异性将大幅降低。 公共卫生相关性：迫切需要新的蛋白质组学技术，以满足新兴的大规模生物学研究的需求。我们已经开发了一种新的微阵列分析平台，它可以使用MALDI TOF质谱成像筛选大珠库。在这里，我们建议将MSBeadArray技术应用于珠缀合肽的文库的开发，所述珠缀合肽将以微阵列形式通过质谱法测量。