Ion mobility and gas phase H/D exchange: revealing the importance of a single hydrogen bond for the chiral recognition of crown ether ammonium complexes

2018 ◽  
Vol 54 (39) ◽  
pp. 4967-4970 ◽  
Author(s):  
Jan M. Wollschläger ◽  
Konstantin Simon ◽  
Marius Gaedke ◽  
Christoph A. Schalley

Ion mobility MS and gas phase H/D exchange experiments allow the detection of chiral recognition of crown ether/ammonium complexes in the gas phase.

The Analyst ◽  
2016 ◽  
Vol 141 (19) ◽  
pp. 5502-5510 ◽  
Author(s):  
Melanie Göth ◽  
Frederik Lermyte ◽  
Xiao Jakob Schmitt ◽  
Stephan Warnke ◽  
Gert von Helden ◽  
...  

The influence of side chain to backbone interactions on the gas-phase structure of ubiquitin and ubiquitin lysine-to-arginine mutants was analysed.


2015 ◽  
Vol 119 (44) ◽  
pp. 10988-10998 ◽  
Author(s):  
Kristian H. Møller ◽  
Anne S. Hansen ◽  
Henrik G. Kjaergaard

2021 ◽  
Vol 14 (6) ◽  
pp. 498
Author(s):  
Evolène Deslignière ◽  
Anthony Ehkirch ◽  
Bastiaan L. Duivelshof ◽  
Hanna Toftevall ◽  
Jonathan Sjögren ◽  
...  

Antibody-drug conjugates (ADCs) are biotherapeutics consisting of a tumor-targeting monoclonal antibody (mAb) linked covalently to a cytotoxic drug. Early generation ADCs were predominantly obtained through non-selective conjugation methods based on lysine and cysteine residues, resulting in heterogeneous populations with varying drug-to-antibody ratios (DAR). Site-specific conjugation is one of the current challenges in ADC development, allowing for controlled conjugation and production of homogeneous ADCs. We report here the characterization of a site-specific DAR2 ADC generated with the GlyCLICK three-step process, which involves glycan-based enzymatic remodeling and click chemistry, using state-of-the-art native mass spectrometry (nMS) methods. The conjugation process was monitored with size exclusion chromatography coupled to nMS (SEC-nMS), which offered a straightforward identification and quantification of all reaction products, providing a direct snapshot of the ADC homogeneity. Benefits of SEC-nMS were further demonstrated for forced degradation studies, for which fragments generated upon thermal stress were clearly identified, with no deconjugation of the drug linker observed for the T-GlyGLICK-DM1 ADC. Lastly, innovative ion mobility-based collision-induced unfolding (CIU) approaches were used to assess the gas-phase behavior of compounds along the conjugation process, highlighting an increased resistance of the mAb against gas-phase unfolding upon drug conjugation. Altogether, these state-of-the-art nMS methods represent innovative approaches to investigate drug loading and distribution of last generation ADCs, their evolution during the bioconjugation process and their impact on gas-phase stabilities. We envision nMS and CIU methods to improve the conformational characterization of next generation-empowered mAb-derived products such as engineered nanobodies, bispecific ADCs or immunocytokines.


Author(s):  
Charles Eldrid ◽  
Aisha Ben-Younis ◽  
Jakub Ujma ◽  
Hannah Britt ◽  
Tristan Cragnolini ◽  
...  

PLoS ONE ◽  
2014 ◽  
Vol 9 (6) ◽  
pp. e100200 ◽  
Author(s):  
Ewa Sitkiewicz ◽  
Jacek Olędzki ◽  
Jarosław Poznański ◽  
Michał Dadlez

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