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

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Jeffrey W Hudgens, Elyssia S Gallagher, Ioannis Karageorgos, Kyle W Anderson, James J Filliben, Richard Y-C Huang, Guodong Chen, George M Bou-Assaf, Alfonso Espada, Michael J Chalmers, Eduardo Harguindey, Hui-Min Zhang, Benjamin T Walters, Jennifer Zhang, John D Venable, Caitlin Steckler, Inhee Park, Ansgar Brock, Xiaojun Lu, Ratnesh K Pandey & 17 andre Arun Chandramohan, Ganesh S Anand, Sasidhar N Nirudodhi, Justin B Sperry, Jason C Rouse, James A Carroll, Kasper D Rand, Ulrike Leurs, David D Weis, Mohammed A Al-Naqshabandi, Tyler S Hageman, Daniel Deredge, Patrick L Wintrode, Malvina Papanastasiou, John D Lambris, Sheng Li, Sarah Urata

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 fifteen 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 〈 sLab 〉 ≤ (0.15 ± 0.01) Da (1σx ̅ ), and all laboratories achieved 〈 sLab 〉 ≤ 0.4 Da. For immersions of protein at THDX = (3.6 to 25) oC and for D2O exchange times of tHDX = (30 s to 4 h) the reproducibility of back-exchange corrected, deuterium uptake measurements for the 15 laboratories is σreproducibility15 Labs ( tHDX ) = (9.0 ± 0.9) % (1σ). A 9 laboratory cohort that immersed samples at THDX = 25 oC exhibited reproducibility of σreproducibility25C cohort ( tHDX ) = (6.5 ± 0.6) % for back-exchange corrected, deuterium uptake measurements.

OriginalsprogEngelsk
TidsskriftAnalytical Chemistry
Vol/bind91
Udgave nummer11
Sider (fra-til)7336-7345
ISSN0003-2700
DOI
StatusUdgivet - 2019

ID: 217934034