Automated Hydroxyl Radical Protein Footprinting for the Determination of Biopharmaceutical Protein Conformation

摘要

A need exists for reliable and sensitive methods to determine higher order structure of pharmaceutical proteins at the onset of biosimilar R&D and regulatory approval. Automation and integration of sub-systems (dosimetry, laser power measurement, enzymology) will lead to increased reproducibility for HRPF based structural characterization. A MRM based method provides a sensitive approach to determine oxidation levels during HRPF. Top down approaches will enable the analysis of conformationally heterogeneous mixtures by HRPF. Ongoing work is focused on applied these methodologies to a suite of pharmaceutical proteins.