Automated Electrophoresis Platform to Streamline Validations of Biomedical Samples

摘要 通用微全分析系统的发展代表了测量的顶峰 科学将样品制备、分析和检测的所有方面集成到一个便宜的， 自动化平台将简化分析，使生物医学样本能够快速验证。的 理想的生物样品分析方法不仅可以验证生物样品的化学成分，而且可以表征 更高级的生物分子结构（例如二硫键、手性、硫酸化）以评估生物活性。到目前为止， 但这些梦想并没有实现。因此，研究人员必须手动制备样品， 用于表征样品纯度的分析，但生物活性的评估往往被忽视。 这种耗时的、不完整的样本验证有可能使后续研究的结果产生偏差。 为了帮助提高NIH资助项目的严谨性和可重复性，我们建议开发一个 通用的生物医学样本验证系统，为研究人员提供快速验证生物医学样本的工具，包括 对决定活性的高级生物结构的评估。热凝胶电泳（TGE） 将成为南极洲的心脏我们的团队开发了TGE来富集，分离和检测 生物分子在温度响应性凝胶内，从而整合了分析方法的多个步骤 变成一个便宜的微流体装置。在我们先前工作的基础上，我们建议进一步扩大我们的 实现理想的综合能力。将开发其他表征， 以高选择性筛选蛋白质、肽、RNA和糖的高级结构， 其他技术（例如LC-MS）无法达到的灵敏度。为了简化分析，采样 制备能力将被集成到设备中，以过滤细胞、脱盐样品和标记分析物 用于检测。这种方法将能够直接分析芯片上的生物样品，从而避免了需要 用于用户的外部样品预处理。此外，还将开发无标签检测方案 以进一步加快分析和简化业务限制。总体而言， 这里开发的策略将提供一种方便，廉价的手段， 这是其他技术无法实现的。最终，我们设想我们的基于TGE的CRTAS 该平台将使研究人员能够获得强大的样本验证，这将提高重现性 生物学研究在学术界，政府和工业实验室。

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.