Charge Variant Analysis of Monoclonal Antibodies Using Direct Coupled pH Gradient Cation Exchange Chromatography to High-Resolution Native Mass Spectrometry

Analytical Chemistry
Florian FüsslJonathan Bones

Abstract

Charge variant analysis (CVA) of monoclonal antibodies (mAbs) using cation exchange chromatography is routinely used as a fingerprint of the distribution of posttranslational modifications present on the molecule. Traditional salt or pH based eluents are not suited for direct coupling to mass spectrometry due to nonvolatility or high ionic strength. This makes further analysis complicated when an alteration in the charge variant profile or the emergence of an additional peak is encountered. Here, the use of pH gradient elution using volatile, low ionic strength buffers is reported with direct coupling to high-resolution Orbitrap mass spectrometry. The development of a universal method based on pH elution was explored using a number of mAb drug products. Optimized methods facilitated the separation and identification of charge variants including individual glycoforms of the mAbs investigated using the same buffer system but with tailored gradient slopes. The developed method represents an exciting advance for the characterization of biopharmaceuticals as intact entities through the combination of native charge variant separations with high-resolution native mass spectrometry.

References

Mar 29, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·R J HarrisA B Chen
Oct 13, 2006·Annual Review of Immunology·James N ArnoldRaymond A Dwek
May 12, 2009·Journal of the American Society for Mass Spectrometry·Alexander Makarov, Eduard Denisov
Feb 16, 2010·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Vera TimmDirk Chelius
Jun 29, 2010·Journal of the American Society for Mass Spectrometry·Peran Terrier, D J Douglas
Apr 18, 2012·Drug Testing and Analysis·Nataša Debeljak, Arthur J Sytkowski
Jun 30, 2012·Nature Reviews. Drug Discovery·Steven A BerkowitzGraham B Jones
Oct 16, 2012·Nature Methods·Rebecca J RoseAlbert J R Heck
Jan 15, 2014·Journal of Separation Science·Michael SchmidtChristian Frech
Sep 10, 2014·Analytical Chemistry·Mohammad TalebiPaul R Haddad
Mar 25, 2015·Journal of Pharmaceutical and Biomedical Analysis·Szabolcs FeketeDavy Guillarme
Apr 19, 2015·Journal of Pharmaceutical and Biomedical Analysis·Szabolcs FeketeDavy Guillarme
Nov 14, 2015·Bioanalysis·Koen Sandra, Pat Sandra
Jan 2, 2016·The Journal of Physical Chemistry Letters·Yulian GavrilovYaakov Levy
Nov 3, 2016·Current Pharmaceutical Biotechnology·Roy Jefferis
Mar 1, 2017·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Y LeblancG Chevreux
Apr 13, 2017·Journal of Chromatography. a·Elsa Wagner-RoussetAlain Beck
Sep 30, 2017·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Alexandre GoyonAlain Beck

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Citations

Jan 9, 2019·Analytical and Bioanalytical Chemistry·Bastiaan L DuivelshofValentina D'Atri
Nov 22, 2019·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Simone SchadtChristoph Funk
Aug 17, 2020·Journal of Chromatography. a·Evelin FarsangSzabolcs Fekete
Jun 23, 2021·Current Opinion in Biotechnology·Florian FüsslJonathan Bones
Jul 22, 2021·Food & Function·Wenhua YangIgor A Kaltashov
Jul 16, 2021·Journal of Pharmaceutical and Biomedical Analysis·Li CaoKevin Lan
Sep 1, 2020·Journal of the American Society for Mass Spectrometry·Yuetian YanNing Li
Aug 21, 2021·Chemical Reviews·Sem TamaraAlbert J R Heck
Sep 3, 2021·Chemical Reviews·Amber D Rolland, James S Prell

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