Localized hydration in lyophilized myoglobin by hydrogen-deuterium exchange mass spectrometry. 2. Exchange kinetics.

Molecular Pharmaceutics
Andreas M Sophocleous, Elizabeth M Topp

Abstract

Solid-state hydrogen-deuterium exchange with mass spectrometric analysis (ssHDX) is a promising method for characterizing proteins in amorphous solids. Though analysis of HDX kinetics is informative and well-established in solution, application of these methods to solid samples is complicated by possible heterogeneities in the solid. The studies reported here provide a detailed analysis of the kinetics of hydration and ssHDX for equine myoglobin (Mb) in solid matrices containing sucrose or mannitol. Water sorption was rapid relative to ssHDX, indicating that ssHDX kinetics was not limited by bulk water transport. Deuterium uptake in solids was well-characterized by a biexponential model; values for regression parameters provided insight into differences between the two solid matrices. Analysis of the widths of peptide mass envelopes revealed that, in solution, an apparent EX2 mechanism prevails, consistent with native conformation of the protein. In contrast, in mannitol-containing samples, a smaller non-native subpopulation exchanges by an EX1-like mechanism. Together, the results indicate that the analysis of ssHDX kinetic data and of the widths of peptide mass envelopes is useful in screening solid formulations of protein dr...Continue Reading

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Citations

Feb 12, 2014·Computers & Chemical Engineering·Brock C RoughtonKyle V Camarda
Nov 15, 2014·Analytical Chemistry·Gregory F PirroneJohn R Engen
Apr 6, 2016·Journal of Pharmaceutical Sciences·Lavanya K IyerElizabeth M Topp
Oct 16, 2013·Molecular Pharmaceutics·Lavanya K IyerElizabeth M Topp
Jan 25, 2021·International Journal of Pharmaceutics·Rishabh TukraElizabeth M Topp
Jun 28, 2019·Molecular Pharmaceutics·Rajashekar Kammari, Elizabeth M Topp
Aug 14, 2020·Molecular Pharmaceutics·Rajashekar Kammari, Elizabeth M Topp

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