Intended Impact
This project is designed to improve, test, and validate hydrogen/deuterium exchange mass spectrometry (HDX-MS) metrology for the determination of dynamical properties of therapeutic proteins and glycoproteins.
Objectives
- Explore structure-function relationships between glycan structure and protein folding energies
- Develop HDX-MS for assessments of dynamical comparability among innovator and candidate biosimilar drugs
- Measure the reproducibility of proteolytic fragmentation HDX-MS by conducting interlaboratory comparisons on a Fab protein
- Improve HDX-MS technology for measurements of trans-membrane protein drug targets
- Measure interactions of biological drugs with surfaces and aggregates
- Enhance resolution of the HDX-MS method to the single amide level
Technical Approach
The research project uses HDX-MS to measure the D-for-H exchange rates of the amide groups along the backbone of a protein in D2O solution. These rates indicate the protection factors of the amide groups. The protection factors are characteristic of higher order structural features of proteins (e.g., α-helixes, β-sheets ...) that are stabilized through hydrogen bonding, disulfide bonds, electrostatic interactions, and hydrophobic forces. Since protection factors can change as the protein binds with ligands or undergoes folding, HDX-MS can provide sensitive diagnostic data evidencing structural differences. HDX-MS is a rapidly evolving metrology. Numerous improvements in automation, instrument resolution and accuracy, and new, more powerful software are being introduced.
