SBIR Phase II: A dual mass spectrometry and fluorescence microarray platform for proteomics

这个小企业创新研究（SBIR）第二阶段项目的更广泛的影响/商业潜力将是开发一种分析方法，使生物样品的质谱和荧光测量。 该检测将基于微阵列平台，用于靶向定量蛋白质组学、酶活性检测和组合化学领域的应用。靶向定量蛋白质组学测量多个样品（如细胞系、组织或生物流体）中特定蛋白质生物标志物浓度的变化。在制药工业中，该方法广泛用于评估药物候选化合物的效率以及其潜在毒性。研究人员对定量蛋白质组学的多重方法特别感兴趣，这些方法可以提供有关各种细胞生化网络状态的详细信息。该项目的意义在于新开发的快速分离、纯化和浓缩大量蛋白质组生物标志物的能力，用于通过质谱进行多重定量分析。这将导致分析小型化，以帮助降低药物开发过程的成本。在制药领域之外，该技术可用于临床环境中进行微创诊断。它还可以扩展到基础生物学研究，以研究广泛的重要生物相互作用，包括抗体-抗原，候选药物-药物靶标，酶-底物，受体-配体等。SBIR第二阶段项目旨在开发一种新的生命科学分析技术，称为微珠辅助质谱法（BAMS）。BAMS是一个微阵列平台，可以通过荧光和质谱法进行独立的数据采集。它应用图像细胞术的方法来快速筛选在微型化微孔板（皮滴定板）上的单个威尔斯孔内排列的数千个微珠。这是第一项将质谱法的高通量分析结合到基于珠的生物测定中的技术，目前主要在流式细胞仪上进行测量。双重检测能力在蛋白质组学中特别重要，其中质谱法可用于区分蛋白质同种型和具有不同翻译后修饰模式（例如磷酸化）的蛋白质。该项目将利用质谱成像结合二维和三维荧光成像，并将在MALDI TOF和ESI MS平台上对珠阵列进行互补分析，以实现更大的序列覆盖率。这项技术将为研究人员提供更多的选择，设计定制的多重珠检测比目前可能的。此外，BAMS可用于测量光学编码的珠、质量标签编码的珠，甚至非编码的珠，例如组合肽珠文库。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project will be to develop an analytical assay to enable measurement of biological samples by mass spectrometry and fluorescence. The assay will be based on a microarray platform, for applications in the areas of targeted quantitative proteomics, enzymatic activity assays, and combinatorial chemistry. Targeted quantitative proteomics measures changes in the concentration of a specific protein biomarker across multiple samples, such as cell lines, tissues, or biofluids. In the pharmaceutical industry, this method is widely used to assess efficiency of a drug candidate compound as well as its potential toxicity. Researchers are especially interested in the multiplexed approaches to quantitative proteomics that could provide detailed information on the state of various cellular biochemical networks. The significance of this project is the newly developed capability to rapidly separate, purify, and concentrate a large number of proteomic biomarkers for multiplexed quantitative analysis by mass spectrometry. This will result in the assay miniaturization to help lower the costs of the drug development process. Outside of the pharmaceutical field, the technology can be utilized in the clinical settings for performing minimally invasive diagnostics. It also can be extended to basic biology research to study a wide range of important biological interactions including antibody-antigen, drug candidate-drug target, enzyme-substrate, receptor-ligand, and others. This SBIR Phase II project proposes to develop a new analytical technology for life sciences termed Bead-Assisted Mass Spectrometry (BAMS). BAMS is a microarray platform that enables independent data acquisition by fluorescence and mass spectrometry. It applies the method of image cytometry to rapidly screen thousands of microbeads arrayed inside individual wells on a miniaturized microwell plate (picotiter plate). It is the first technology to incorporate high throughput analysis by mass spectrometry into bead-based bioassays, which in their current form are measured primarily on flow cytometry instruments. The dual detection capability is particularly important in proteomics where mass spectrometry can be applied to distinguish protein isoforms and proteins with different patterns of post-translational modifications such as phosphorylation. This project will utilize mass spectrometry imaging in combination with two- and three-dimensional fluorescence imaging, and will perform complementary analysis of bead arrays on the MALDI TOF and ESI MS platforms to achieve greater sequence coverage. This technology will provide researchers with far more options with respect to designing custom multiplex bead assays than currently possible. In addition, BAMS may be used to measure optically encoded beads, mass tag-encoded beads, and even non-encoded beads such as combinatorial peptide bead libraries.