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

点击翻译按钮获取中文摘要

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.