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

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; 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

doi:10.1021/acs.analchem.9b01100

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

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 PandeyArun 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

Molecular Cardiology and Membrane Proteins

19 Citations (Scopus)

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^Lab»≤ (0.15 ± 0.01) Da (1σ). All laboratories achieved s^Lab»≤ 0.4 Da. For immersions of protein at T_HDX = (3.6 to 25) °C and for D₂O exchange times of t_HDX = (30 s to 4 h) the reproducibility of back-exchange corrected, deuterium uptake measurements for the 15 laboratories is σreproducibility15 Laboratories(t_HDX) = (9.0 ± 0.9) % (1σ). A nine laboratory cohort that immersed samples at T_HDX = 25 °C exhibited reproducibility of σreproducibility25C cohort(t_HDX) = (6.5 ± 0.6) % for back-exchange corrected, deuterium uptake measurements.

Original language	English
Journal	Analytical Chemistry
Volume	91
Issue number	11
Pages (from-to)	7336-7345
ISSN	0003-2700
DOIs	https://doi.org/10.1021/acs.analchem.9b01100
Publication status	Published - 4 Jun 2019

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Hudgens, J. W., Gallagher, E. S., Karageorgos, I., Anderson, K. W., Filliben, J. J., Huang, R. Y-C., Chen, G., Bou-Assaf, G. M., Espada, A., Chalmers, M. J., Harguindey, E., Zhang, H-M., Walters, B. T., Zhang, J., Venable, J. D., Steckler, C., Park, I., Brock, A., Lu, X., ... Urata, S. (2019). Interlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab fragment of NISTmAb. Analytical Chemistry, 91(11), 7336-7345. https://doi.org/10.1021/acs.analchem.9b01100

Hudgens, 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, JD, Steckler, C, Park, I, Brock, A, Lu, X, Pandey, RK, Chandramohan, A, Anand, GS, 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 2019, 'Interlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab fragment of NISTmAb', Analytical Chemistry, vol. 91, no. 11, pp. 7336-7345. https://doi.org/10.1021/acs.analchem.9b01100

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title = "Interlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab fragment of NISTmAb",

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 sLab»≤ (0.15 ± 0.01) Da (1σ). All laboratories achieved sLab»≤ 0.4 Da. For immersions of protein at THDX = (3.6 to 25) °C 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 Laboratories(tHDX) = (9.0 ± 0.9) % (1σ). A nine laboratory cohort that immersed samples at THDX = 25 °C exhibited reproducibility of σreproducibility25C cohort(tHDX) = (6.5 ± 0.6) % for back-exchange corrected, deuterium uptake measurements.",

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

year = "2019",

month = jun,

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doi = "10.1021/acs.analchem.9b01100",

language = "English",

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pages = "7336--7345",

journal = "Industrial And Engineering Chemistry Analytical Edition",

issn = "0003-2700",

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T1 - Interlaboratory Comparison of Hydrogen-Deuterium Exchange Mass Spectrometry Measurements of the Fab fragment of NISTmAb

AU - Hudgens, Jeffrey W

AU - Gallagher, Elyssia S

AU - Karageorgos, Ioannis

AU - Anderson, Kyle W

AU - Filliben, James J

AU - Huang, Richard Y-C

AU - Chen, Guodong

AU - Bou-Assaf, George M

AU - Espada, Alfonso

AU - Chalmers, Michael J

AU - Harguindey, Eduardo

AU - Zhang, Hui-Min

AU - Walters, Benjamin T

AU - Zhang, Jennifer

AU - Venable, John D

AU - Steckler, Caitlin

AU - Park, Inhee

AU - Brock, Ansgar

AU - Lu, Xiaojun

AU - Pandey, Ratnesh K

AU - Chandramohan, Arun

AU - Anand, Ganesh S

AU - Nirudodhi, Sasidhar N

AU - Sperry, Justin B

AU - Rouse, Jason C

AU - Carroll, James A

AU - Rand, Kasper D

AU - Leurs, Ulrike

AU - Weis, David D

AU - Al-Naqshabandi, Mohammed A

AU - Hageman, Tyler S

AU - Deredge, Daniel

AU - Wintrode, Patrick L

AU - Papanastasiou, Malvina

AU - Lambris, John D

AU - Li, Sheng

AU - Urata, Sarah

PY - 2019/6/4

Y1 - 2019/6/4

N2 - 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 sLab»≤ (0.15 ± 0.01) Da (1σ). All laboratories achieved sLab»≤ 0.4 Da. For immersions of protein at THDX = (3.6 to 25) °C 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 Laboratories(tHDX) = (9.0 ± 0.9) % (1σ). A nine laboratory cohort that immersed samples at THDX = 25 °C exhibited reproducibility of σreproducibility25C cohort(tHDX) = (6.5 ± 0.6) % for back-exchange corrected, deuterium uptake measurements.

AB - 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 sLab»≤ (0.15 ± 0.01) Da (1σ). All laboratories achieved sLab»≤ 0.4 Da. For immersions of protein at THDX = (3.6 to 25) °C 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 Laboratories(tHDX) = (9.0 ± 0.9) % (1σ). A nine laboratory cohort that immersed samples at THDX = 25 °C exhibited reproducibility of σreproducibility25C cohort(tHDX) = (6.5 ± 0.6) % for back-exchange corrected, deuterium uptake measurements.

U2 - 10.1021/acs.analchem.9b01100

DO - 10.1021/acs.analchem.9b01100

M3 - Journal article

C2 - 31045344

SN - 0003-2700

VL - 91

SP - 7336

EP - 7345

JO - Industrial And Engineering Chemistry Analytical Edition

JF - Industrial And Engineering Chemistry Analytical Edition

IS - 11

ER -

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

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