Automated Electrophoresis Platform to Streamline Validations of Biomedical Samples

自动化电泳平台可简化生物医学样品的验证

• 批准号: 10710812
• 负责人: Thomas Linz
• 金额: $ 32.84万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-21 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10710812
• 关键词: Biological; Biomedical Research; Cells; Chemicals; Consumption; Detection; Development; Devices; Dreams; Electrophoresis; Evaluation; Gel; Government; Heart; Higher Order Chromatin Structure; Industry; Intervention; Label; Laboratories; Manuals; Measurement; Microfluidic Microchips; Peptides; Preparation; RNA; Reproducibility; Research Personnel; Risk; Sampling; Scheme; Science; Structure; Sulfate; System; Techniques; Temperature; Time; United States National Institutes of Health; Validation; Work; analytical method; disulfide bond; gel electrophoresis; improved; innovation; micro-total analysis system; neglect; protein structure; research study; sugar; tool

ABSTRACT Development of a universal micro total analysis system (TAS) represents the pinnacle of measurement science. Integrating all aspects of sample preparation, analysis, and detection into an inexpensive, automated platform would streamline analyses and enable rapid validation of biomedical samples. The ideal TAS would not only validate the chemical composition of a biological sample, but also characterize higher order biomolecule structure (e.g. disulfide bonds, chirality, sulfation) to evaluate bioactivity. To date, though, these dreams have not been realized. Consequently, researchers must manually prepare samples for analyses that characterize sample purity, but assessments of biological activity often remain neglected. This time-consuming, incomplete sample validation risks biasing results of subsequent research studies. To help improve the rigor and reproducibility of NIH-sponsored projects, we propose to develop a universal TAS to provide researchers with a tool to rapidly validate biomedical samples, including evaluations of higher order biological structures that dictate activity. Thermal gel electrophoresis (TGE) will serve as the heart of the TAS. Our group developed TGE to enrich, separate, and detect biomolecules within a temperature-responsive gel, thus integrating multiple steps of an analytical method into an inexpensive microfluidic device. Building on our prior work, we propose to further expand our capabilities towards the ideal comprehensive TAS. Additional characterizations will be developed to screen the higher order structure of proteins, peptides, RNAs, and sugars with high selectivity and sensitivity that are inaccessible to other techniques (e.g. LC-MS). To streamline analyses, sample preparation capabilities will be integrated into devices to filter cells, desalt samples, and label analytes for detection. This approach will enable direct analysis of biological samples on-chip, obviating the need for external sample pretreatment by the user. Additionally, label-free detection schemes will be developed to further expedite analyses and simplify operational constraints. Collectively, the innovative analytical strategies developed here will provide a convenient, inexpensive means of characterizing biomedical samples that cannot be achieved by other techniques. Ultimately, we envision our TGE-based TAS platform will make robust sample validation accessible to researchers, which will increase reproducibility of biological studies in academic, government, and industry laboratories.

抽象的 通用微型总分析系统（TA）的开发代表测量的巅峰 科学。将样品制备，分析和检测的所有方面整合到廉价的， 自动化平台将简化分析并能够快速验证生物医学样本。这 理想的tas不仅会验证生物样品的化学成分，而且还表征 高阶生物分子结构（例如二硫键，手性，硫化）来评估生物活性。迄今为止， 虽然，这些梦想尚未实现。因此，研究人员必须手动准备样本 对于表征样品纯度的分析，但对生物活性的评估通常仍被忽略。 这项耗时的，不完整的样本验证风险随后研究的偏见结果。 为了帮助提高NIH赞助项目的严格性和可重复性，我们建议开发一个 通用tas为研究人员提供一种工具，以快速验证生物医学样本，包括 对决定活动的高阶生物结构的评估。热凝胶电泳（TGE） 将充当the tas的心脏。我们的小组开发了TGE来丰富，分开和检测 温度响应性凝胶中的生物分子，从而整合了分析方法的多个步骤 进入廉价的微流体设备。在我们先前的工作的基础上，我们建议进一步扩大我们的 理想综合tas的能力。将开发其他字符 筛选具有高选择性的蛋白质，肽，RNA和糖的高阶结构 其他技术（例如LC-MS）无法访问的灵敏度。为了简化分析，样品 制备功能将集成到设备中，以进行滤清器，删除样品和标签分析物 用于检测。这种方法将可以直接分析片上的生物样品，从而消除了需求 用于用户的外部样本预处理。此外，将制定无标签检测方案 进一步加快分析并简化操作约束。总体而言，创新的分析 这里开发的策略将提供一种方便，廉价的方式来表征生物医学 其他技术无法实现的样品。最终，我们设想了基于TGE的塔 平台将使研究人员可以访问强大的样本验证，这将增加可重复性 学术，政府和行业实验室的生物学研究。