Integrated Digital Microfluidic Platforms for Next-Generation Glycomics

摘要 蛋白质糖基化广泛存在于生命的各个领域，是阐明分子生物学的关键 支撑疾病的机制。尽管葡聚糖具有生物医学和临床意义，但在这方面的研究进展 由于巨大的结构多样性，Glycomics大大落后于基因组学和蛋白质组学 以及由复杂的非模板驱动的生物合成产生的葡聚糖的动态变化。因此，就是这样 必须开发创新的技术和仪器，以促进在 血糖分析。这项提案旨在解决下一代糖组分的迫切需求 通过利用来自PI实验室研究的新微流控平台，开发了一种新型微流控技术。 具体地说，PI建议开发一种新的基于微流控执行器的单分子分析方法 这将大大提高凝集素微阵列的分析性能。此外，私家侦探寻求 将样本处理功能与数字凝集素-抗体条形码阵列系统集成在一起，构建了 灵敏定量靶向血糖“样品-输入-输出”数字微流控凝集素阵列平台 以高通量的方式进行分析。该项目包括三个目标：1)研制数字微流控芯片 低丰度糖蛋白的单分子定量方法；2)建立完全整合的 用于多重、定量靶向糖链分析的微系统；以及3)展示定量 卵巢癌患者血浆的比较血糖谱。这项研究如果成功，将产生一种 系统糖组学的关键变革工具，并使PI能够追求生物医学的长期利益 通过获取人体动态概况研究蛋白质糖基化的病理作用 血浆糖蛋白。

Abstract Protein glycosylation is ubiquitously involved in all domains of life and holds the key to elucidating molecular mechanisms underpinning disease. Despite the biomedical and clinical significance of glycans, progress in glycomics has considerably lagged behind genomics and proteomics, owing to enormous structural diversity and dynamic alternation of glycans arising from complex non-template driven biosynthesis. Thus it is imperative to develop innovative technologies and instrumentation that can catalyze key breakthroughs in glycomic analysis. This proposal intends to address the urgent need of the next-generation glycomics technologies by leveraging on new microfluidic platforms stemming from research in the PI's laboratory. Specifically, the PI proposes to develop a novel microfluidic actuator-based single-molecule analysis method that can substantially improve the analytical performance of lectin microarray. Moreover, the PI seeks to integrate sample processing functionalities with the digital lectin-antibody barcode array system to construct a “sample-in-answer-out” digital microfluidic lectin array platform for sensitive and quantitative targeted glycomic profiling in a high-throughput manner. The project consists of three aims: 1) develop a digital microfluidic approach for single-molecule quantification of low-abundance glycoproteins; 2) develop a fully integrated microsystem for multiplexed, quantitative targeted glycomic profiling; and 3) demonstrate quantitative comparative glycomic profiling of plasma from ovarian cancer patients. This research, if successful, will yield a key transformative tool for systems glycomics and allow the PI to pursue the long-term interest in biomedical investigation of pathological roles of protein glycosylation through obtaining dynamic overview of human plasma glycome.