Automated Electrophoresis Platform to Streamline Validations of Biomedical Samples

摘要 通用微量全分析系统(-TAS)的开发代表着测量的顶峰 科学。将样品制备、分析和检测的所有方面集成到一个廉价的、 自动化平台将简化分析并实现生物医学样本的快速验证。这个 理想的原子吸收光谱不仅可以验证生物样品的化学成分，还可以表征 高阶生物分子结构(如二硫键、手性、硫酸盐化)以评价生物活性。到目前为止， 然而，这些梦想还没有实现。因此，研究人员必须手动准备样本 用于表征样品纯度的分析，但对生物活性的评估往往被忽视。 这种耗时的、不完整的样本验证可能会对后续研究的结果产生偏差。 为了帮助提高NIH赞助的项目的严密性和重复性，我们建议开发一种 UniversalTAS为研究人员提供快速验证生物医学样本的工具，包括 对决定活动的高级生物结构的评估。热凝胶电泳(TGE) 将作为TAS的心脏。我们团队开发了TGE来丰富、分离和检测 温度响应凝胶内的生物分子，从而集成了分析方法的多个步骤 变成一种廉价的微流控装置。在我们先前工作的基础上，我们建议进一步扩大我们的 向理想的全面TAS迈进的能力。将开发更多的特征描述以 筛选蛋白质、肽、RNA和糖的高级结构，具有高选择性和 其他技术(如LC-MS)无法获得的灵敏度。要简化分析，请采样 准备能力将集成到设备中，以过滤细胞、脱盐样本和标记分析物 用来侦测。这种方法将能够在芯片上直接分析生物样本，从而消除了 供用户进行外部样品前处理。此外，还将开发无标签检测方案 以进一步加快分析和简化操作限制。总体而言，创新的分析 这里开发的策略将提供一种方便、廉价的手段来表征生物医学 其他技术无法实现的样品。最终，我们设想我们基于TGE的TAS 平台将使研究人员能够访问健壮的样本验证，这将增加重复性 学术、政府和工业实验室的生物学研究。

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.