Structural Analysis of Intact Monoclonal Antibodies by Electron Transfer Dissociation Mass Spectrometry

Tsybin, Yury O.; Fornelli, Luca; Stoermer, Carsten; Luebeck, Markus; Parra, Julien; Nallet, Sophie; Wurm, Florian Μ.; Hartmer, Ralf

doi:10.1021/ac201293m

Cited by 132 publications

(143 citation statements)

References 54 publications

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“…Antibodies have also been analyzed using this instrument, allowing for improved sequence coverage through the use of electron transfer dissociation (ETD) of the disulfide intact species [85]. ETD is an electron-based fragmentation technique similar to ECD, but utilizes gaseous anions to transfer low-energy electrons to protonated analytes [86].…”

Section: Orbitrap Mass Spectrometrymentioning

confidence: 99%

Top-down proteomics: applications, recent developments and perspectives

Pamreddy

Panyala

2016

J Appl Bioanal

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REVIEWNow-a-days, top-down proteomics (TDP) is a booming approach for the analysis of intact proteins and it is attaining significant interest in the field of protein biology. The term has emerged as an alternative to the well-established, bottom-up strategies for analysis of peptide fragments derived from either enzymatically or chemically digestion of intact proteins. TDP is applied to mass spectrometric analysis of intact large biomolecules that are constituents of protein complexes and assemblies. This article delivers an overview of the methodologies in top-down mass spectrometry, mass spectrometry instrumentation and an extensive review of applications covering the venomics, biomedical research, protein biology including the analysis of protein post-translational modifications (PTMs), protein biophysics, and protein complexes. In addition, limitations of top-down proteomics, challenges and future directions of TDP are also discussed.

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Section: Orbitrap Mass Spectrometrymentioning

confidence: 99%

Top-down proteomics: applications, recent developments and perspectives

Pamreddy

Panyala

2016

J Appl Bioanal

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“…Topdown proteomics and petroleomics, because of their extreme requirements for analytical performance, are the primary application areas of FT-ICR MS nowadays [3,7]. Over the past decade, other application areas of interest were primarily addressed using Orbitrap FTMS and high-resolution time-offlight (TOF) MS [4,[8][9][10][11]. Nevertheless, the extreme analytical challenges in molecular structural analysis require further increase of FTMS performance.…”

Section: Introductionmentioning

confidence: 99%

Ion Trap with Narrow Aperture Detection Electrodes for Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Nagornov

Kozhinov

Tsybin

et al. 2015

J. Am. Soc. Mass Spectrom.

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Abstract. The current paradigm in ion trap (cell) design for Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) is the ion detection with wide aperture detection electrodes. Specifically, excitation and detection electrodes are typically 90°wide and positioned radially at a similar distance from the ICR cell axis. Here, we demonstrate that ion detection with narrow aperture detection electrodes (NADEL) positioned radially inward of the cell's axis is feasible and advantageous for FT-ICR MS. We describe design details and performance characteristics of a 10 T FT-ICR MS equipped with a NADEL ICR cell having a pair of narrow aperture (flat) detection electrodes and a pair of standard 90°excitation electrodes. Despite a smaller surface area of the detection electrodes, the sensitivity of the NADEL ICR cell is not reduced attributable to improved excite field distribution, reduced capacitance of the detection electrodes, and their closer positioning to the orbits of excited ions. The performance characteristics of the NADEL ICR cell are comparable with the state-of-the-art FT-ICR MS implementations for small molecule, peptide, protein, and petroleomics analyses. In addition, the NADEL ICR cell's design improves the flexibility of ICR cells and facilitates implementation of advanced capabilities (e.g., quadrupolar ion detection for improved mainstream applications). It also creates an intriguing opportunity for addressing the major bottleneck in FTMS-increasing its throughput via simultaneous acquisition of multiple transients or via generation of periodic non-sinusoidal transient signals.

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“…It also preserves the labile modifications that are likely destroyed by bottom-up MS and introduce minimal artificial modifications, which result from lengthy sample preparation processes such as overnight digestion. These distinct features make topdown MS ideal for analyzing protein pharmaceuticals such as monoclonal antibodies (Bondarenko et al 2009, Tsybin et al 2011. For disulfide bond assignment, top-down MS can be useful because it avoids possible disulfide rearrangement that usually occurs in mild alkaline conditions during trypsin digestion.…”

Section: Introductionmentioning

confidence: 99%

Mass spectrometry-based strategies for protein disulfide bond identification

Tsai

Chen²,

Huang

2013

Reviews in Analytical Chemistry

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Abstract:The formation of disulfide bonds is critical for stabilizing protein structures and maintaining protein functions. It is important to understand the linkages between multiple cysteine residues within a protein. In this review, the analytical approaches using mass spectrometry (MS) for disulfide linkage assignment are classified and discussed. Enzymatic digestion under appropriate conditions followed by various MS detection strategies remains the primary method for cysteine linkage analysis. In-source decay (ISD) and electron transfer dissociation (ETD) have been used to generate significant peptide signals that indicate the identities of peptides involved in disulfide bonds. In addition, chemical labeling and software algorithms were also developed to facilitate the automation of disulfide bond analysis. For proteins with complex disulfide structure, methods involving partial reduction coupled with differential alkylation were demonstrated to be useful. In the past two decades, MS has become one of the most valuable tools for protein disulfide bond analysis. It provides irreplaceable information including the peptide backbone sequences as well as the cysteine connection pattern when coupling with appropriate sample preparations. The related approaches with their unique features can be applied for different aims such as structural characterization or functional studies of proteins.

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Structural Analysis of Intact Monoclonal Antibodies by Electron Transfer Dissociation Mass Spectrometry

Cited by 132 publications

References 54 publications

Top-down proteomics: applications, recent developments and perspectives

Top-down proteomics: applications, recent developments and perspectives

Ion Trap with Narrow Aperture Detection Electrodes for Fourier Transform Ion Cyclotron Resonance Mass Spectrometry

Mass spectrometry-based strategies for protein disulfide bond identification

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