We describe a set of simple devices for surface-induced dissociation proteins and protein complexes on three instrument platforms. All the use novel yet split lens geometry that is minimally invasive (requiring few millimeters along ion path axis) easier to operate than prior generations devices. The designed be small enough replace entrance Bruker FT-ICR collision cell, dynamic range enhancement (DRE) Waters Q-IM-TOF, or exit transfer multipole Thermo Scientific Extended Mass Range (EMR)...

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...

Mass-spectrometry based assays in structural biology studies measure either intact or digested proteins. Typically, different mass spectrometers are dedicated for such measurements: those optimized rapid analysis of peptides designed high molecular weight analysis. A commercial trapped ion mobility-quadrupole-time-of-flight (TIMS-Q-TOF) platform is widely utilized proteomics and metabolomics, with mobility providing a separation dimension addition to liquid chromatography. The ability...

"Top-down" proteomics analyzes intact proteins and identifies proteoforms by their mass as well the observed fragmentation pattern in tandem spectrometry (MS/MS) experiments. Recently, hybrid ion mobility spectrometry–mass (IM/MS) methods have gained traction for top-down experiments, either allowing analysis of individual isomers or alternatively improving signal/noise dynamic range fragment assignment. We recently described construction a tandem-trapped spectrometer/mass spectrometer...

A second-generation ("Gen 2") device capable of surface-induced dissociation (SID) and collision-induced (CID) for Fourier transform ion cyclotron resonance mass spectrometry protein complexes has been designed, simulated, fabricated, experimentally compared to a first-generation 1"). The primary goals the redesign were (1) simplify SID by reducing number electrodes, (2) increase CID sensitivity lengthening collision cell, (3) range analysis larger multimeric proteins, all while maintaining...

Native mass spectrometry (nMS), particularly in conjunction with gas-phase ion mobility measurements, has proven useful as a structural biology tool for evaluating the stoichiometry, conformation, and topology of protein complexes. Here, we demonstrate combination trapped (TIMS) surface-induced dissociation (SID) on Bruker SolariX XR 15 T FT-ICR spectrometer analysis We successfully performed SID mobility-selected complexes, including streptavidin tetramer cholera toxin B bound ligands....

Mass-spectrometry based assays in structural biology studies measure either intact or digested proteins. Typically, different mass spectrometers are dedicated for such measurements: those optimized rapid analysis of peptides designed high molecular weight analysis. A commercial trapped ion mobility-quadrupole-time flight (TIMS-Q-TOF) platform is widely utilized proteomics and metabolomics, with mobility providing a separation dimension addition to liquid chromatography. The ability perform...

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 spectrometry)-Quadrupole-SID (surface-induced dissociation)-Time Flight, TIMS-Q-SID-TOF, instrument nMS. The modifications include changing cartridge from...

The use of submicrometer capillaries for nanoelectrospray ionization native proteins and protein complexes effectively reduces the number nonspecific salt adducts to biological molecules, therefore increasing apparent resolution a mass spectrometer without any further instrument modifications or increased ion activation. However, interaction between surface capillary has been shown promote expansion loss structure. Here, we compare effect micrometer sized on structures streptavidin,...

The complexity of the proteome at all levels protein structure and dynamics renders systematic identification how modifications alter protein-protein interaction networks cause disease a formidable, unsolved challenge. Mass spectrometry-methods enable detection causing on proteome-wide scale but do not provide direct structural information to reveal molecular origins disease-causing events. Here, we demonstrate that (i) ultraviolet photodissociation coupled with tandem-trapped ion mobility...

Mass-spectrometry based assays in structural biology studies measure either intact or digested proteins. Typically, different mass spectrometers are dedicated for such measurements: those optimized rapid analysis of peptides designed high molecular weight analysis. A commercial trapped ion mobility-quadrupole-time flight (TIMS-Q-TOF) platform is widely utilized proteomics and metabolomics, with mobility providing a separation dimension addition to liquid chromatography. The ability perform...

Investigating the structural heterogeneity of monoclonal antibodies is crucial to achieving optimal therapeutic outcomes. We show that tandem-trapped ion mobility spectrometry enables collision-induced unfolding measurements antibody subpopulations. Our data suggest such non-ensemble will improve characterisation complex biotherapeutics as multivalent antibodies.

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...

We describe a set of simple devices for surface-induced dissociation protein complexes on three instrument platforms. All the use novel yet split lens geometry that is minimally invasive (requiring few mm along ion path axis) and easier to operate than prior generations devices. The designed be small enough replace entrance Bruker FT-ICR collision cell, dynamic range enhancement (DRE) Waters Q-IM-TOF, or exit transfer multipole Thermo Scientific Extended Mass Range (EMR) Orbitrap. used SID...

<p><a>Native mass spectrometry, particularly in conjunction with gas-phase ion mobility spectrometry measurements, has proven useful as a structural biology tool for evaluating the stoichiometry, conformation, and topology of protein complexes. Here, we demonstrate combination trapped (TIMS) surface-induced dissociation (SID) on Bruker SolariX XR 15 T FT-ICR spectrometer analysis We successfully performed SID mobility-selected complexes, including streptavidin tetramer cholera...

Mass spectrometry-based assays in structural biology studies typically measure either intact or digested proteins. Traditionally, there are different mass spectrometers dedicated for such measurements: those focused on the rapid analysis of small molecules, as peptides, and designed high analysis. The Bruker timsTOF Pro spectrometer (ion mobility-quadrupole-time flight platform, IM-Q-TOF), has become widely utilized molecules fields proteomics metabolomics, with ion mobility spectrometry...

Investigating the structural heterogeneity of monoclonal antibodies is crucial to achieving optimal therapeutic outcomes. We show that tandem-trapped ion mobility spectrometry enables collision-induced unfolding measurements antibody subpopulations. Our data suggest such non-ensemble will improve characterisation complex biotherapeutics as multivalent antibodies.

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 spectrometry)-Quadrupole-SID (surface-induced dissociation)-Time Flight, TIMS-Q-SID-TOF, instrument nMS. The modifications include changing cartridge from...

The amino acid position within a histone sequence and the chemical nature of post-translational modifications (PTMs) are essential for elucidating "Histone Code". Previous work has shown that PTMs induce specific biological responses good candidates as biomarkers diagnostics. Here, we evaluate analytical advantages trapped ion mobility (TIMS) with parallel accumulation-serial fragmentation (PASEF) tandem mass spectrometry (MS/MS) bottom-up proteomics model cancer cells. study also considered...

Investigating the structural heterogeneity of monoclonal antibodies is crucial to achieving optimal therapeutic outcomes. We show that tandem-trapped ion mobility spectrometry enables collision-induced unfolding measurements subpopulations a humanised IgGk NIST antibody (NISTmAb). Our results indicate differential glycosylation NISTmAb does not modulate its conformational heterogeneity.

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 spectrometry)-Quadrupole-SID (surface-induced dissociation)-Time Flight, TIMS-Q-SID-TOF, instrument nMS. The modifications include changing cartridge from...

Mass spectrometry-based assays in structural biology studies typically measure either intact or digested proteins. Traditionally, there are different mass spectrometers dedicated for such measurements: those focused on the rapid analysis of small molecules, as peptides, and designed high analysis. The Bruker timsTOF Pro spectrometer (ion mobility-quadrupole-time flight platform, IM-Q-TOF), has become widely utilized molecules fields proteomics metabolomics, with ion mobility spectrometry...

<p>We describe a set of simple devices for surface-induced dissociation protein complexes on three instrument platforms. All the use novel yet split lens geometry that is minimally invasive (requiring few mm along ion path axis) and easier to operate than prior generations devices. The designed be small enough replace entrance Bruker FT-ICR collision cell, dynamic range enhancement (DRE) Waters Q-IM-TOF, or exit transfer multipole Thermo Scientific Extended Mass Range (EMR) Orbitrap....

Native mass spectrometry, particularly in conjunction with gas-phase ion mobility spectrometry measurements, has proven useful as a structural biology tool for evaluating the stoichiometry, conformation, and topology of protein complexes. Here, we demonstrate combination trapped (TIMS) surface-induced dissociation (SID) on Bruker SolariX XR 15 T FT-ICR spectrometer analysis We successfully performed SID mobility-selected complexes, including streptavidin tetramer cholera toxin B bound...