NMR-compatible BioMEMS platform for in vitro and in vivo protein structure and function characterization

• Applicants: Dr. Vlad Badilita; Dr. Neil MacKinnon, Ph.D.
• Subject Area: Microsystems
• Term: from 2016 to 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 268738192

The BIO-PRICE goal is to address a key challenge of modern molecular biology: development of a robust tool to allow NMR investigation of proteins at work in a living environment. Our first aim is to develop a BioMEMS environmental platform enabling on-demand protein and cell immobilization with high spatial and temporal resolution at electro-addressable locations inside the platform. Next, we plan to integrate carefully designed NMR µ-detectors for high-resolution investigation of the immobilized target at the previously defined locations. Merging microsystem- and bio-engineering will enable us to create a network of electro-addressable investigation sites and to reconstitute the living environment so that, in the final stage, integration of µNMR detectors will address the issue of high-throughput protein NMR investigation. We have identified several research objectives which need to be addressed so that the results will constitute a useful tool for both NMR and biosystems research communities.O1: Replicate the natural environment of proteins for the purpose of NMR protein investigation. The objectives pertaining to ensuring a living environment, which replicates the natural environment where proteins operate, address both in vitro and in vivo (in-cell) investigation.O2: On-demand protein loading and immobilization with high spatial resolution inside the NMR detection volume for in vitro investigation. In order to be able to perform statistically relevant and/or combinatorial measurement datasets, fast and automatic loading and positioning of the protein at the most favorable position from the point of view of NMR sensitivity and field uniformity inside the µNMR detection volume is required.O3: Develop a dedicated platform for in-cell protein NMR. Microfluidic devices for in-cell protein NMR must provide sufficient aeration, cell-nurturing media supplied to the measurement site, accumulated waste and metabolites timely evacuated.O4: Simultaneous detection of multiple protein-relevant nuclear species for multi-dimensional, hetero-nuclear measurements - development of miniaturized NMR detectors capable of targeting up to 3 different Larmor frequencies, so that the same detector can be used to capture information from multiple nuclei of interest at the same time.O5: Address long acquisition times by increasing the detector sensitivity through miniaturization. Another objective is material engineering at detector level towards obtaining zero-susceptibility, low resistivity sensor material (e.g., wire) for minimum B0-field distortion, maximum signal pickup and low noise.O6: Enable high-throughput, parallel protein NMR investigation. The development of a platform capable of interrogating multiple nuclear species in a broad spectral range would open the way for a wide range of applications ranging from metabolomics and protein-medical drug screening, to low-cost targeted applications such as point of care investigation or low field NMR.

• DFG Programme: Research Grants