Improving mass accuracy of high performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry of intact antibodies.

Journal of the American Society for Mass Spectrometry
Himanshu S GadgilPavel V Bondarenko

Abstract

The glycosylation profile of intact antibody due to the galactose and fucose heterogeneity in the N-linked sugars was determined with instrument resolution of 5000 and 10,000. After deconvolution of electrospray ionization mass spectra to complete convergence, several extra peaks appeared in addition to the peaks observed in the original mass spectra. The artificial peaks were avoided if deconvolution was stopped after a smaller number of iterations. A standard antibody was used as an external calibrant to minimize mass measurement errors during long-period experiments. Precision of four consecutive LC/MS measurements of the same antibody was 10 ppm (+/-1.5 Da). By using this approach, the masses of 11 intact antibodies were measured. All antibodies containing N-terminal glutamines had a negative mass shift due to the formation of pyroglutamate (-17 Da). Although the pyroglutamate variant of intact antibody was not resolved from the unmodified variant, this modification led to a mass shift proportional to the percentage of N-terminal pyroglutamate. By accurately measuring the mass shift we were able to quantify the abundance of pyroglutamic acid on intact antibodies. Mass accuracy in measuring different antibodies was below 30 ...Continue Reading

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Citations

Oct 15, 2013·Analytical and Bioanalytical Chemistry·Natalia NavasLuis Cuadros-Rodríguez
Feb 9, 2008·Journal of the American Society for Mass Spectrometry·Lowell J BradyAlain Balland
Aug 19, 2008·Journal of the American Society for Mass Spectrometry·Sandipan SinhaHimanshu S Gadgil
May 5, 2009·Journal of the American Society for Mass Spectrometry·Pavel V BondarenkoZhongqi Zhang
Nov 24, 2011·BMC Biotechnology·Yoan J MachadoMartin Himly
Feb 3, 2011·Journal of Pharmaceutical Sciences·Amanda K MillerHimanshu S Gadgil
Aug 6, 2008·Journal of Pharmaceutical Sciences·Nicholas Van BurenJaby Jacob
Jul 26, 2011·Journal of Pharmaceutical Sciences·Steven D StroopScott D Leigh
Jul 27, 2011·Journal of Pharmaceutical Sciences·Nicole StackhouseHimanshu S Gadgil
Jul 11, 2007·Journal of Pharmaceutical Sciences·Himanshu S GadgilMichael Treuheit
Dec 17, 2014·Biological & Pharmaceutical Bulletin·Yoichi IshiiKaori Wakamatsu
Jul 24, 2012·Drug Discovery Today·Jingjie MoGuodong Chen
Aug 23, 2008·Mass Spectrometry Reviews·Zhongqi ZhangXiaoyu Chen
Feb 13, 2009·Proteomics·Carolin HuhnManfred Wuhrer
Nov 28, 2009·Journal of Mass Spectrometry : JMS·Jennifer ZhangViswanatham Katta
Mar 20, 2014·Vaccine·K de la Luz-HernándezA Castillo
Aug 1, 2014·Bioanalysis·Chris A Singleton
Nov 14, 2006·Biotechnology and Bioengineering·Lawrence W DickKuang-Chuan Cheng
Jul 3, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·Alain BeckNathalie Corvaïa
Oct 30, 2019·Proceedings of the National Academy of Sciences of the United States of America·Nicholas A MoringoChristy F Landes
Jun 18, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Unyong KimSang Beom Han
Feb 28, 2007·Journal of Mass Spectrometry : JMS
Jul 14, 2009·Analytical Chemistry·Pavlo PristatskyJosef Vlasak
Nov 8, 2007·Analytical Chemistry·Lowell J BradyAlain Balland
Nov 2, 2019·Journal of Pharmaceutical and Biomedical Analysis·Julien CamperiNathalie Delaunay
Apr 23, 2021·Journal of the American Society for Mass Spectrometry·Iain D G Campuzano, Wendy Sandoval

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