Interlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab Fragment of NISTmAb.

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TitleInterlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab Fragment of NISTmAb.
Publication TypeJournal Article
Year of Publication2019
AuthorsHudgens, JW, Gallagher, ES, Karageorgos, I, Anderson, KW, Filliben, JJ, Huang, RY-C, Chen, G, Bou-Assaf, GM, Espada, A, Chalmers, MJ, Harguindey, E, Zhang, H-M, Walters, BT, Zhang, J, Venable, J, Steckler, C, Park, I, Brock, A, Lu, X, Pandey, R, Chandramohan, A, Anand, GSrinivasan, Nirudodhi, SN, Sperry, JB, Rouse, JC, Carroll, JA, Rand, KD, Leurs, U, Weis, DD, Al-Naqshabandi, MA, Hageman, TS, Deredge, D, Wintrode, PL, Papanastasiou, M, Lambris, JD, Li, S, Urata, S
JournalAnal Chem
Date Published2019 May 14
ISSN1520-6882
Abstract

Hydrogen-deuterium exchange mass spectrometry (HDX-MS) is an established, powerful tool for investigating protein-ligand interactions, protein folding, and protein dynamics. However, HDX-MS is still an emergent tool for quality control of biopharmaceuticals and for establishing dynamic similarity between a biosimilar and an innovator therapeutic. Because industry will conduct quality control and similarity measurements over a product lifetime and in multiple locations, an understanding of HDX-MS reproducibility is critical. To determine the reproducibility of continuous-labeling, bottom-up HDX-MS measurements, the present interlaboratory comparison project evaluated deuterium uptake data from the Fab fragment of NISTmAb reference material (PDB: 5K8A ) from 15 laboratories. Laboratories reported ∼89 800 centroid measurements for 430 proteolytic peptide sequences of the Fab fragment (∼78 900 centroids), giving ∼100% coverage, and ∼10 900 centroid measurements for 77 peptide sequences of the Fc fragment. Nearly half of peptide sequences are unique to the reporting laboratory, and only two sequences are reported by all laboratories. The majority of the laboratories (87%) exhibited centroid mass laboratory repeatability precisions of ⟨ s⟩ ≤ (0.15 ± 0.01) Da (1σ). All laboratories achieved ⟨s⟩ ≤ 0.4 Da. For immersions of protein at T = (3.6 to 25) °C and for DO exchange times of t = (30 s to 4 h) the reproducibility of back-exchange corrected, deuterium uptake measurements for the 15 laboratories is σ( t) = (9.0 ± 0.9) % (1σ). A nine laboratory cohort that immersed samples at T = 25 °C exhibited reproducibility of σ( t) = (6.5 ± 0.6) % for back-exchange corrected, deuterium uptake measurements.

DOI10.1021/acs.analchem.9b01100
Alternate JournalAnal. Chem.
PubMed ID31045344