A simple cellulose column procedure for selective enrichment of glycopeptides and characterization by nano LC coupled with electron-transfer and high-energy collisional-dissociation tandem mass spectrometry

2010 ◽  
Vol 345 (6) ◽  
pp. 792-801 ◽  
Author(s):  
Sergei I. Snovida ◽  
Edward D. Bodnar ◽  
Rosa Viner ◽  
Julian Saba ◽  
Hélène Perreault
Keyword(s):  
Mass Spectrometry ◽  
Electron Transfer ◽  
Tandem Mass Spectrometry ◽  
High Energy ◽  
Tandem Mass ◽  
Selective Enrichment ◽  
Cellulose Column ◽  
Column Procedure

scholarly journals Structural Characterization of Daunomycin-Peptide Conjugates by Various Tandem Mass Spectrometric Techniques

2021 ◽  
Vol 22 (4) ◽  
pp. 1648
Author(s):  
Adina Borbély ◽  
Lilla Pethő ◽  
Ildikó Szabó ◽  
Mohammed Al-Majidi ◽  
Arnold Steckel ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Electron Transfer ◽  
Tandem Mass Spectrometry ◽  
High Energy ◽  
Amino Acid Sequences ◽  
Tandem Mass ◽  
Peptide Conjugates ◽  
Maldi Tof ◽  
Tandem Mass Spectrometric

The use of peptide-drug conjugates has generated wide interest as targeted antitumor therapeutics. The anthracycline antibiotic, daunomycin, is a widely used anticancer agent and it is often conjugated to different tumor homing peptides. However, comprehensive analytical characterization of these conjugates via tandem mass spectrometry (MS/MS) is challenging due to the lability of the O-glycosidic bond and the appearance of MS/MS fragment ions with little structural information. Therefore, we aimed to investigate the optimal fragmentation conditions that suppress the prevalent dissociation of the anthracycline drug and provide good sequence coverage. In this study, we comprehensively compared the performance of common fragmentation techniques, such as higher energy collisional dissociation (HCD), electron transfer dissociation (ETD), electron-transfer higher energy collisional dissociation (EThcD) and matrix-assisted laser desorption/ionization–tandem time-of-flight (MALDI-TOF/TOF) activation methods for the structural identification of synthetic daunomycin-peptide conjugates by high-resolution tandem mass spectrometry. Our results showed that peptide backbone fragmentation was inhibited by applying electron-based dissociation methods to conjugates, most possibly due to the “electron predator” effect of the daunomycin. We found that efficient HCD fragmentation was largely influenced by several factors, such as amino acid sequences, charge states and HCD energy. High energy HCD and MALDI-TOF/TOF combined with collision induced dissociation (CID) mode are the methods of choice to unambiguously assign the sequence, localize different conjugation sites and differentiate conjugate isomers.

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