Adapting a trapped ion mobility spectrometry-Q-TOF for native mass spectrometry DOI Creative Commons
Yu-Fu Lin,

Benjamin Jones,

Mark E. Ridgeway

et al.

Published: May 9, 2024

Native mass spectrometry (nMS) is increasingly popular for studying intact protein quaternary structure. When coupled with ion mobility, which separates ions based on their size, charge, and shape, it provides additional structural information the complex of interest. In this study, we present a novel prototype TIMS (trapped mobility)-Quadrupole-SID (surface-induced dissociation)-Time Flight (TIMS-Q-SID-TOF) instrument nMS. The modifications include changing cartridge from concave to convex geometry electrodes operating at 425 kHz improve trapping efficiency high mass-to-charge (m/z) mobility analysis, such as 3 4 MDa hepatitis B virus capsids. quadrupole radiofrequency driver was lowered 385 kHz, extends isolation range 3,000 17,000 m/z allows single charge state GroEL 16,200 an window 25 m/z. Finally, 6-mm thick, 2-lens SID device installed replaced collision cell entrance lens. dissociated 801 kDa into all combinations subcomplexes, peaks were well-resolved easy interpret. This first time timsTOF Pro nMS has been introduced resolving power separation selection fragmentation product collection across broad 1,500 40,000.

Language: Английский

Native Mass Spectrometry: Recent Progress and Remaining Challenges DOI
Kelly R. Karch, Dalton T. Snyder, Sophie R. Harvey

et al.

Annual Review of Biophysics, Journal Year: 2022, Volume and Issue: 51(1), P. 157 - 179

Published: Jan. 4, 2022

Native mass spectrometry (nMS) has emerged as an important tool in studying the structure and function of macromolecules their complexes gas phase. In this review, we cover recent advances nMS related techniques including sample preparation, instrumentation, activation methods, data analysis software. These have enabled nMS-based to address a variety challenging questions structural biology. The second half review highlights applications these technologies surveys classes that can be studied with nMS. Complementarity existing biology current challenges are also addressed.

Language: Английский

Citations

99
Top-down proteomics DOI
David S. Roberts, Joseph A. Loo, Yury O. Tsybin

et al.

Nature Reviews Methods Primers, Journal Year: 2024, Volume and Issue: 4(1)

Published: June 13, 2024

Language: Английский

Citations

20
Native top‐down mass spectrometry for higher‐order structural characterization of proteins and complexes DOI
Ruijie Liu,

Shujun Xia,

Huilin Li

et al.

Mass Spectrometry Reviews, Journal Year: 2022, Volume and Issue: 42(5), P. 1876 - 1926

Published: June 27, 2022

Abstract Progress in structural biology research has led to a high demand for powerful and yet complementary analytical tools characterization of proteins protein complexes. This significantly increased interest native mass spectrometry (nMS), particularly top‐down (nTDMS) the past decade. review highlights recent advances nTDMS biological assemblies, with particular focus on extra multi‐layers information enabled by TDMS. We include short introduction sample preparation ionization nMS, tandem fragmentation techniques as well analyzers software/analysis pipelines used nTDMS. highlight unique offered examples its broad range applications proteins, protein‐ligand interactions (metal, cofactor/drug, DNA/RNA, protein), therapeutic antibodies antigen‐antibody complexes, membrane macromolecular machineries (ribosome, nucleosome, proteosome, viruses), endogenous The challenges, potential, along perspectives methods analysis assemblies recombinant samples are discussed.

Language: Английский

Citations

40
Approaches to Heterogeneity in Native Mass Spectrometry DOI
Amber D. Rolland, James S. Prell

Chemical Reviews, Journal Year: 2021, Volume and Issue: 122(8), P. 7909 - 7951

Published: Sept. 2, 2021

Native mass spectrometry (MS) is aimed at preserving and determining the native structure, composition, stoichiometry of biomolecules their complexes from solution after they are transferred into gas phase. Major improvements in MS instrumentation experimental methods over past few decades have led to a concomitant increase complexity heterogeneity samples that can be analyzed, including protein–ligand complexes, protein with multiple coexisting stoichiometries, membrane protein–lipid assemblies. Heterogeneous features these biomolecular important for understanding structure function. However, sample make assignment ion mass, charge, very challenging due overlap tens or even hundreds peaks spectrum. In this review, we cover data analysis, experimental, instrumental advances strategies solving problem, an in-depth discussion theoretical practical aspects use available deconvolution algorithms tools. We also reflect upon current challenges provide view future exciting field.

Language: Английский

Citations

55
N-Glycosylation of monoclonal antibody therapeutics: A comprehensive review on significance and characterization DOI

Anuj Shrivastava,

Srishti Joshi, András Guttman

et al.

Analytica Chimica Acta, Journal Year: 2022, Volume and Issue: 1209, P. 339828 - 339828

Published: April 13, 2022

Language: Английский

Citations

36
Online Hydrophilic Interaction Chromatography (HILIC) Enhanced Top-Down Mass Spectrometry Characterization of the SARS-CoV-2 Spike Receptor-Binding Domain DOI
Jesse Wilson, Aivett Bilbao, Juan Wang

et al.

Analytical Chemistry, Journal Year: 2022, Volume and Issue: 94(15), P. 5909 - 5917

Published: April 5, 2022

SARS-CoV-2 cellular infection is mediated by the heavily glycosylated spike protein. Recombinant versions of protein and receptor-binding domain (RBD) are necessary for seropositivity assays can potentially serve as vaccines against viral infection. RBD plays key roles in protein's structure function, thus, comprehensive characterization recombinant critically important biopharmaceutical applications. Liquid chromatography coupled to mass spectrometry has been widely used characterize post-translational modifications proteins, including glycosylation. Most studies RBDs were performed at proteolytic peptide (bottom-up proteomics) or released glycan level because technical challenges resolving highly heterogeneous glycans intact level. Herein, we evaluated several online separation techniques: (1) C2 reverse-phase liquid (RPLC), (2) capillary zone electrophoresis (CZE), (3) acrylamide-based monolithic hydrophilic interaction (HILIC) separate with varying combinations glycosylations (glycoforms) top-down (MS). Within conditions explored, HILIC method was superior RPLC CZE separating glycoforms, which differ significantly neutral groups. In addition, our analysis readily captured unexpected (e.g., cysteinylation N-terminal sequence variation) low abundance, proteoforms that may be missed using glycopeptide data alone. The MS platform holds great potential glycoproteins facile comparison biosimilars quality control

Language: Английский

Citations

23
Development of in silico models to predict viscosity and mouse clearance using a comprehensive analytical data set collected on 83 scaffold-consistent monoclonal antibodies DOI Creative Commons
Marissa Mock,

Alex W. Jacobitz,

Christopher J. Langmead

et al.

mAbs, Journal Year: 2023, Volume and Issue: 15(1)

Published: Sept. 12, 2023

Biologic drug discovery pipelines are designed to deliver protein therapeutics that have exquisite functional potency and selectivity while also manifesting biophysical characteristics suitable for manufacturing, storage, convenient administration patients. The ability use computational methods predict properties from sequence, potentially in combination with high throughput assays, could decrease timelines increase the success rates therapeutic developability engineering by eliminating lengthy expensive cycles of recombinant production testing. To support development high-quality predictive models antibody developability, we a sequence-diverse panel 83 effector functionless IgG1 antibodies displaying range properties, produced formulated each under standard platform conditions, collected comprehensive package analytical data, including vitro assays vivo mouse pharmacokinetics. We used this robust training data set build machine learning classifier can complex behavior these features derived predicted and/or experimental structures. Our 87% accuracy whether viscosity at 150 mg/mL is above or below threshold 15 centipoise (cP) 75% area plasma concentration-time curve (AUC0-672 h) normal 3.9 × 106 h x ng/mL.

Language: Английский

Citations

16
Fragment-based drug discovery campaigns guided by native mass spectrometry DOI
Louise M. Sternicki, Sally‐Ann Poulsen

RSC Medicinal Chemistry, Journal Year: 2024, Volume and Issue: 15(7), P. 2270 - 2285

Published: Jan. 1, 2024

Native mass spectrometry (nMS) is well established as a biophysical technique for characterising biomolecules and their interactions with endogenous or investigational small molecule ligands such fragments.

Language: Английский

Citations

4
Standardized workflow for multiplexed charge detection mass spectrometry on orbitrap analyzers DOI
Pei Su, John P. McGee, Michael A. R. Hollas

et al.

Nature Protocols, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 2, 2025

Language: Английский

Citations

0
Adapting a Trapped Ion Mobility Spectrometry-Q-TOF for High m/z Native Mass Spectrometry and Surface-Induced Dissociation DOI
Yu-Fu Lin, Benjamin J. Jones, Mark E. Ridgeway

et al.

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 16, 2025

Native mass spectrometry (nMS) is an increasingly popular technique for studying intact protein quaternary structure. When coupled with ion mobility, which separates ions based on their size, charge, and shape, it provides additional structural information the complex of interest. We present here data from a novel prototype TIMS (trapped mobility spectrometry)-quadrupole-SID (surface-induced dissociation)-time flight, TIMS-Q-SID-TOF, instrument nMS. The modifications include changing cartridge concave to convex electrode geometry dual tunnel design operating at 425 kHz radio frequency (RF) improve trapping efficiency high mass-to-charge (m/z) analysis, such as 3 4 MDa hepatitis B virus capsids. quadrupole RF driver was lowered 385 kHz, extends isolation range 3,000 17,000 m/z allows single charge state GroEL 16,200 window 25 m/z. Finally, 6 mm thick, 2-lens SID device replaced collision cell entrance lens. dissociated 801 kDa into all combinations subcomplexes, peaks were well-resolved, allowing confident assignment product ions. This first time timsTOF Pro nMS has been introduced resolving power separation selection fragmentation collection detection across broad 1,500 40,000.

Language: Английский

Citations

0