PEPTIDE BEAD MICROARRAYS MEASURED BY MASS SPECTROMETRY IMAGING

描述（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。我们开发了一种新型的微阵列分析平台，可以使用MALDI TOF质谱成像筛选大型头部库。虽然通过质谱法进行生物组织成像现在是一项成熟的技术，但MALDI TOF MSI尚未用于进行生物微阵列扫描。我们的MSBeadArray技术能够在一定条件下将选定的分析物从微珠转移到特殊设计的微阵列载玻片表面，从而保持分析物与单个微珠的空间分离，同时保持从同一微珠中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个样品的头库进行质谱分析，双荧光- MSI读出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技术应用于头部共轭肽文库的开发，这些文库将通过微阵列格式的质谱法进行测量。与具有质谱读数的肽头阵列相比，具有位置编码的平面肽微阵列和具有荧光编码的悬浮头阵列（例如，Luminex“平台）具有显著的优势。由于质谱法可以在一次测量中检测到多种肽，因此在微阵列扫描期间可以测量肽的纯度，附加序列的存在，甚至每个肽的相对数量。肽制造商通常使用MALDI TOF质谱分析对用于生产微阵列的合成肽进行质量控制。然而，制备的微阵列没有被质谱分析，因此固定在微阵列载玻片或微球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头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，头测定的可变性将大大减少。

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.