Sampling strategies for capillary isoelectric focusing with electroosmotic zone mobilization assessed by high-resolution dynamic computer simulation

2012 ◽  
Vol 33 (6) ◽  
pp. 970-980 ◽  
Author(s):  
Anna Takácsi-Nagy ◽  
Ferenc Kilár ◽  
Csilla Páger ◽  
Richard A. Mosher ◽  
Wolfgang Thormann
Keyword(s):  
Computer Simulation ◽  
High Resolution ◽  
Isoelectric Focusing ◽  
Capillary Isoelectric Focusing ◽  
Sampling Strategies ◽  
Dynamic Computer Simulation

An improved capillary isoelectric focusing-mass spectrometry method for high-resolution characterization of monoclonal antibody charge variants

2022 ◽  
Author(s):  
Tian Xu ◽  
Linjie Han ◽  
Alayna M. George Thompson ◽  
Liangliang Sun
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
High Resolution ◽  
Isoelectric Focusing ◽  
Capillary Isoelectric Focusing ◽  
Spectrometry Method ◽  
Mass Spectrometry Method ◽  
Charge Variants

Routine and high-resolution characterization of monoclonal antibody (mAb) charge variants is vital for controlling mAb quality as therapeutics. Capillary isoelectric focusing-mass spectrometry (cIEF-MS) has emerged as a powerful tool for...

scholarly journals A Mini Review on Capillary Isoelectric Focusing-Mass Spectrometry for Top-Down Proteomics

2021 ◽  
Vol 9 ◽  
Author(s):  
Tian Xu ◽  
Liangliang Sun
Keyword(s):  
Mass Spectrometry ◽  
High Resolution ◽  
Isoelectric Focusing ◽  
Large Scale ◽  
Protein Characterization ◽  
Capillary Isoelectric Focusing ◽  
Top Down ◽  
Esi Ms ◽  
Highly Sensitive

Mass spectrometry (MS)-based top-down proteomics (TDP) requires high-resolution separation of proteoforms before electrospray ionization (ESI)-MS and tandem mass spectrometry (MS/MS). Capillary isoelectric focusing (cIEF)-ESI-MS and MS/MS could be an ideal method for TDP because cIEF can enable separation of proteoforms based on their isoelectric points (pIs) with ultra-high resolution. cIEF-ESI-MS has been well-recognized for protein characterization since 1990s. However, the widespread adoption of cIEF-MS for the characterization of proteoforms had been impeded by several technical challenges, including the lack of highly sensitive and robust ESI interface for coupling cIEF to MS, ESI suppression of analytes from ampholytes, and the requirement of manual operations. In this mini review, we summarize the technical improvements of cIEF-ESI-MS for characterizing proteoforms and highlight some recent applications to hydrophobic proteins, urinary albumin variants, charge variants of monoclonal antibodies, and large-scale TDP of complex proteomes.

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