CAREER: Developing Thermal Gel Electrophoresis to Interrogate Higher Order Biological Structure

在化学系化学测量和成像计划的支持下，以及化学、生物工程、环境和运输系统系的部分共同资助下，韦恩州立大学的Thomas Linz博士和他的团队正在开发测量生物分子的创新方法。蛋白质必须在生物样本和药物配方中具有“活性”，才能发挥治疗功能。为了提高生物研究研究的重现性和筛选生物制药产品的完整性，需要简便、低成本的方法来测定样品中的活性蛋白质含量，而不是目标蛋白质的总量。林茨实验室正在通过开发技术来快速确定处于活性状态的蛋白质分子的比例，以满足这一需求。一种名为热凝胶电泳的技术正在被创造出来，它使用对温度敏感的材料来帮助区分活性和非活性蛋白质分子。在林茨实验室创建的方法旨在快速和廉价地分析生物样本和生物制药。该小组还为中小学生开发了相关的推广活动，以教孩子们温度如何影响材料的性质。蛋白质的生物活性在一定程度上受折叠构象、多聚体复合体和翻译后修饰的支配。在验证生物样品或生物制药配方时，对这些高阶结构的筛选可以帮助确定和控制样品中活性蛋白质的数量，而不仅仅是总蛋白质质量。林茨实验室正在开发微芯片凝胶电泳格式的方法，以方便地询问三级和四级结构以及翻译后修饰。热凝胶电泳使用热敏聚合物在单个微流控设备中集成不同的分析功能，包括样品准备、分离和检测。这些不同的功能是通过以高空间和时间分辨率控制设备中的温度来实现的，以局部调整每个分析步骤的凝胶粘度。这种方法将微型设备的复杂性和成本降至最低，并简化了分析以增加样品吞吐量。最终，这种快速、廉价的凝胶电泳筛选技术有望提高生物研究的精确度，并使蛋白质药物和疫苗配方的完整性能够在现场进行评估。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, and partial co-funding from the Division of Chemical, Bioengineering, Environmental, and Transport Systems, Dr. Thomas Linz and his group at Wayne State University are developing innovative methods to measure biological molecules. Proteins must be “active” in biological samples and pharmaceutical formulations to carry out therapeutic functions. To improve the reproducibility of biological research studies and to screen the integrity of biopharmaceutical products, convenient low-cost methods are needed for determination of the amount of active protein in a sample, as opposed to the total amount of a target protein. The Linz Lab is addressing this need by developing technology to rapidly determine the fraction of protein molecules in their active states. A technique called thermal gel electrophoresis is being created that uses temperature-responsive materials to help distinguish active and inactive protein molecules. The methods created in the Linz Lab target rapid and inexpensive analysis of biological samples and biopharmaceuticals. The group is also developing associated outreach activities for elementary and middle school students to teach children how temperature impacts the properties of materials.Protein bioactivity is governed in part by folded conformations, multimeric complexes, and post-translational modifications. Screening for these higher order structures when validating biological samples or biopharmaceutical formulations can help determine and control the amount of active protein in the sample, rather than just total protein mass. The Linz Lab is developing methods in a microchip gel electrophoresis format to conveniently interrogate tertiary and quaternary structures and post-translational modifications. Thermal gel electrophoresis uses thermoresponsive polymers to integrate distinct analytical capabilities within a single microfluidic device including sample preparation, separation, and detection. These disparate functions are achieved by controlling temperature in the device with high spatial and temporal resolution to locally adjust gel viscosity for each analysis step. This approach minimizes complexity and cost of the microdevices and streamlines analyses to increase sample throughput. Ultimately, this rapid, inexpensive gel electrophoresis screening technique is expected to enhance the precision of biological research and enable the integrity of protein drug and vaccine formulations to be assessed in the field.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.