Development of High-throughput Multidimensional Collision Induced Unfolding Technology

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor Brandon Ruotolo and his group at the University of Michigan are developing powerful new tools for characterizing the three-dimensional structures of proteins and RNA – both key classes of biomacromolecules in life processes. Specifically, the Ruotolo group is expanding the capabilities of "collision induced unfolding" (CIU), an approach that studies protein structures by quickly removing the water which surrounds biological molecules in their normal environment, then induces their unfolding while monitoring their size and shape using a state-of-the-art tool known as ion mobility-mass spectrometry (IM-MS).

The approach has the potential to revolutionize the detail and acquisition speed with which biomolecular stability can be evaluated. Potential applications include the rapid analysis of next-generation engineered biotherapeutics. The work will provide excellent training for students interested in research at the interface between chemical measurements and the pharmaceutical sciences.

Under this award, the Ruotolo research team at the University of Michigan aims to addresses three main research goals. Firstly, the team will work on building improved methods for the operation of cyclic-type IM-MS instruments for improved retention of native-like biomolecular structure, collision cross-section (CCS) calibration accuracy, and higher-order IM experiments (e.g., IM-CIU and CIUN).

Secondly, and in parallel, the Michigan team will endeavor to develop next-generation software that incorporates features for automated peak picking, feature skipping, on-board CCS calibration, and support for CIUN data types. Then Professor Ruotolo and his students aim to leverage these improved IM-MS and CIU tools to study antibodies and large RNAs in an effort to build assays that support cutting-edge biomolecular engineering campaigns of significance to the biotechnology sector.

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.