Implementation of a design of experiments to study the influence of the background electrolyte on separation and detection in non-aqueous capillary electrophoresis-mass spectrometry

2012 ◽  
Vol 33 (4) ◽  
pp. 583-598 ◽  
Author(s):  
Tjorben Nils Posch ◽  
Alexander Müller ◽  
Wolfgang Schulz ◽  
Michael Pütz ◽  
Carolin Huhn
2018 ◽  
Vol 25 (3) ◽  
pp. 324-332 ◽  
Author(s):  
Jérémie Giorgetti ◽  
Antony Lechner ◽  
Elise Del Nero ◽  
Alain Beck ◽  
Yannis-Nicolas François ◽  
...  

Capillary electrophoresis-mass spectrometry coupling is a growing technique in biopharmaceutics characterization. Assessment of monoclonal antibodies is well known at middle-up and bottom-up levels to obtain information about the sequence, post-translational modifications and degradation products. Intact protein analysis is an actual challenge to be closer to the real protein structure. At this level, actual techniques are time consuming or cumbersome processes. In this work, a 20 minutes separation method has been developed to optimize characterization of intact monoclonal antibodies. Thus, separation has been done on a positively charged coated capillary with optimized volatile background electrolyte and sample buffer. Three world-wide health authorities approved monoclonal antibodies have been used to set up a rapid and ease of use method. Intact trastuzumab, rituximab and palivizumab isoforms have been partially separated with this method in less than 20 minutes under denaturing conditions. For each monoclonal antibody, 2X-glycosylated and 1X-glycosylated structures have been identified and separated. Concerning basic and acidic variants, potential aspartic acid isomerization modification and asparagine deamidation have been observed. Accurate mass determination for high-mass molecular species remains a challenge, but the progress in intact monoclonal antibodies separation appears very promising for biopharmaceutics characterization.


1998 ◽  
Vol 817 (1-2) ◽  
pp. 49-57 ◽  
Author(s):  
Yoshihide Tanaka ◽  
Yasuhiro Kishimoto ◽  
Koji Otsuka ◽  
Shigeru Terabe

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