scholarly journals Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry

iScience ◽  
2021 ◽  
pp. 103211
Author(s):  
Szu-Hsueh Lai ◽  
Sem Tamara ◽  
Albert J.R. Heck
Keyword(s):  
Mass Spectrometry ◽  
Single Particle ◽  
Particle Mass ◽  
Mass Analysis ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection

scholarly journals Probing Affinity, Avidity, Anti-Cooperativity, and Competition in Antibody and Receptor Binding to the SARS-CoV-2 Spike by Single Particle Mass Analyses

2021 ◽  
Author(s):  
Victor C Yin ◽  
Szu-Hsueh Lai ◽  
Tom G Caniels ◽  
Philip J.M. Brouwer ◽  
Mitch Brinkkemper ◽  
...  
Keyword(s):  
Receptor Binding ◽  
Single Particle ◽  
Binding Sites ◽  
Particle Mass ◽  
Mass Analysis ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection ◽  
Analysis Techniques ◽  
Complex Interactions ◽  
Igg Binding

Determining how antibodies interact with the spike (S) protein of the SARS-CoV-2 virus is critical for combating COVID-19. Structural studies typically employ simplified, truncated constructs that may not fully recapitulate the behaviour of the original complexes. Here, we combine two single particle mass analysis techniques (mass photometry and charge-detection mass spectrometry) to enable measurement of full IgG binding to the trimeric SARS-CoV-2 S ectodomain. Our experiments reveal that antibodies targeting the S-trimer typically prefer stoichiometries lower than the symmetry-predicted 3:1 binding. We determine that this behaviour arises from the interplay of steric clashes and avidity effects that are not reflected in common antibody constructs (i.e. Fabs). Surprisingly, these sub-stoichiometric complexes are fully effective at blocking ACE2 binding despite containing free receptor binding sites. Our results highlight the importance of studying antibody/antigen interactions using complete, multimeric constructs and showcase the utility of single particle mass analyses in unraveling these complex interactions.

scholarly journals Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry

2019 ◽  
Author(s):  
Tobias P. Wörner ◽  
Joost Snijder ◽  
Antonette Bennett ◽  
Mavis Agbandje-McKenna ◽  
Alexander A. Makarov ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Single Particle ◽  
High Mass ◽  
Measured Signal ◽  
Particle Charge ◽  
Protein Assemblies ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection ◽  
Adeno Associated Virus ◽  
Virus Particles

AbstractHere we show that single particle charge-detection mass spectrometry (CD-MS) can be performed on a ubiquitous Orbitrap mass analyser and applied to the analysis of high-mass (megadalton) heterogeneous biomolecular assemblies. We demonstrate that single particle high-mass ions can survive in the Orbitrap for seconds, whereby their measured signal amplitudes scale linearly with charge state over the entire m/z range. Orbitrap based single particle CD-MS can be used to resolve mixed ion populations, accurately predict charge states, and consequently also the mass of the ions. We successfully applied CD-MS to challenging natural and biotherapeutic protein assemblies, such as IgM oligomers, designed protein nano-cages, ribosome particles and intact, empty- and genome-loaded Adeno-associated virus particles. Single particle CD-MS combined with native MS on existing Orbitrap platforms will greatly expand its application, especially in the mass analysis of megadalton heterogeneous biomolecular assemblies.

Resolving heterogeneous macromolecular assemblies by Orbitrap-based single-particle charge detection mass spectrometry

2020 ◽  
Vol 17 (4) ◽  
pp. 395-398 ◽  
Author(s):  
Tobias P. Wörner ◽  
Joost Snijder ◽  
Antonette Bennett ◽  
Mavis Agbandje-McKenna ◽  
Alexander A. Makarov ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Single Particle ◽  
Particle Charge ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection ◽  
Macromolecular Assemblies

Characterization of vertical aerosol flows by single particle mass spectrometry for micrometeorological analysis

2011 ◽  
Vol 102 (1-2) ◽  
pp. 49-56 ◽  
Author(s):  
Elmar Gelhausen ◽  
Klaus-Peter Hinz ◽  
Andres Schmidt ◽  
Bernhard Spengler
Keyword(s):  
Mass Spectrometry ◽  
Single Particle ◽  
Particle Mass ◽  
Single Particle Mass Spectrometry

Heterogeneity of Glycan Processing on Trimeric SARS-CoV-2 Spike Protein Revealed by Charge Detection Mass Spectrometry

2021 ◽  
Vol 143 (10) ◽  
pp. 3959-3966 ◽  
Author(s):  
Lohra M. Miller ◽  
Lauren F. Barnes ◽  
Shannon A. Raab ◽  
Benjamin E. Draper ◽  
Tarick J. El-Baba ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Spike Protein ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection

Probing Antibody Binding to Canine Parvovirus with Charge Detection Mass Spectrometry

2018 ◽  
Vol 140 (46) ◽  
pp. 15701-15711 ◽  
Author(s):  
Carmen A. Dunbar ◽  
Heather M. Callaway ◽  
Colin R. Parrish ◽  
Martin F. Jarrold
Keyword(s):  
Mass Spectrometry ◽  
Canine Parvovirus ◽  
Antibody Binding ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection

Characterization of Classical Vaccines by Charge Detection Mass Spectrometry

2021 ◽  
Vol 93 (35) ◽  
pp. 11965-11972 ◽  
Author(s):  
Lohra M. Miller ◽  
Kevin M. Bond ◽  
Benjamin E. Draper ◽  
Martin F. Jarrold
Keyword(s):  
Mass Spectrometry ◽  
Charge Detection Mass Spectrometry ◽  
Charge Detection
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