Dr. Frank Delaglio
Friday February 23, 2018
10:00am, CIPR 120
"Fingerprinting the High Order Structure of Biologic Therapeutics with NMR"
Growth of biologic therapeutics is outpacing that of small-molecule drugs, and the development, manufacture, and delivery of biologics presents very different challenges. Among biologics, monoclonal antibodies (mAbs) have become particularly attractive, since it is possible select and duplicate antibodies that bind with high affinity and specificity to most any target, and biomanufacturing, platforms for mAbs can be reused for more than one therapeutic. Characterization of biologics and their formulations requires monitoring high order structure (HOS), since misfolding or aggregation can lead to loss of efficacy or cause unintended and potentially life-threatening immune responses. Nuclear Magnetic Resonance (NMR), which can provide detailed information on structure and dynamics at atomic resolution, is a powerful tool to probe HOS. That said, heteronuclear NMR carries its own challenges: typical biomolecular NMR applications use isotopically enriched samples, long measurement times, and need extensive and often subjective interactive analysis by an expert in order to extract structural information. We show that fast NMR measurement techniques combined with Non-Uniform Sampling (NUS) strategies can generate practical two-dimensional 1H/13C spectra at natural isotopic abundance for molecules as large as intact monoclonal antibodies. Using measurements on the IgG1k NIST reference mAb (NISTmAb), we demonstrate that small variations of structure and interaction can be revealed and classified by direct computational analysis of the shapes of such spectra, as an alternative to interactive analysis and assignment of spectral features. This paves the way for NMR characterization of biologics via chemometrics and machine learning that is both objective and automated.
~Coffee/tea will be served prior to lecture.~