PEPTIDE BEAD MICROARRAYS MEASURED BY MASS SPECTROMETRY IMAGING

通过质谱成像测量肽珠微阵列

• 批准号: 8546431
• 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 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8546431
• 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; 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; screening; 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.

描述（由申请人提供）：迫切需要新的蛋白质组学技术，以满足新出现的大规模生物学研究的需求。我们已经开发了一个新型的微阵列分析平台，该平台可以使用MALDI TOF质谱成像对大型珠库进行筛选。虽然质谱法的生物组织成像现在是一种完善的技术，但MALDI TOF MSI尚未用于对生物微阵列进行扫描。我们的MSBEADARRAY技术使所选分析物从珠子中转移到条件下特殊设计的微阵列幻灯片的表面，该条件保持了从单个微粒的空间分离，同时保留从同一珠子中提取的分析物的共定位分析物。该平台的独特功能包括对单个微阵列幻灯片上多达500,000个样品的珠库的MS分析，双荧光 - MSI读数以及执行无标签样品检测的选项，以消除对荧光标签和检测抗体的需求。在这里，我们建议将MSBEADARRAY技术应用于珠偶联肽库的开发，该肽将通过微阵列格式通过质谱测量。具有质谱读数的肽珠阵列将比两个平面肽微阵列具有显着优势微阵列扫描。由于MSBEADARRAY平台提供的探针质量数据，微阵列反应将大大降低珠测定的变异性。