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EP38454
Poster Title: N and O Glycosylation in Recombinant SARS CoV 2 RBD Expression Products Analyzed by Next Generation MALDI Top Down Sequencing
Submitted on 09 Mar 2022
Author(s): Arndt Asperger 1 ; Anja Resemann 1 ; Waltraut Evers 1 ; Niels Goedecke 1 ; Ryan Marsico 2 ; Detlev Suckau 1
Affiliations: 1 Bruker Daltonics GmbH & Co. KG, Bremen, Germany; 2 Bruker Scientific LLC, Billerica, MA
Poster Views: 245
Submitted on 09 Mar 2022
Author(s): Arndt Asperger 1 ; Anja Resemann 1 ; Waltraut Evers 1 ; Niels Goedecke 1 ; Ryan Marsico 2 ; Detlev Suckau 1
Affiliations: 1 Bruker Daltonics GmbH & Co. KG, Bremen, Germany; 2 Bruker Scientific LLC, Billerica, MA
Poster Views: 245
Abstract: The receptor binding domain (RBD) of SARS CoV 2 S glycoprotein plays a key role in the interaction with the ACE2 receptor on host cells and is, therefore, of interest for the development of rapid COVID19 antigen tests. As protein modifications are affecting protein function, RBD expression products need to be characterized in depth with regard to their N and O glycosylation profile.
We describe here a novel method for rapid Top Down analysis of N and O glycosylation in recombinant SARS CoV 2 RBDs utilizing Next Generation MALDI Top Down Sequencing (Next Gen MALDI TDS). Next Gen MALDI TDS combines MALDI In Source Decay (ISD) with ultrafast separation by Trapped Ion Mobility Spectrometry (TIMS), and high resolution, accurate time of flight mass analysis (TOF MS), delivering information simultaneously regarding primary sequence, terminal status and near terminal modifications of intact proteins.Summary: We describe here a novel method for rapid Top Down analysis of N and O glycosylation in recombinant SARS CoV 2 RBDs utilizing Next Generation MALDI Top Down Sequencing (Next Gen MALDI TDS).References: [1] C. Gstöttner, T. Zhang, A. Resemann, S. Ruben, S. Pengelley, D. Suckau, T. Welsink, M. Wuhrer, E. Domínguez Vega. Anal. Chem. 2021, 93, 17, 6839-6847
[2] A. Asperger, A. Resemann, W. Evers, N. Goedecke, D. Suckau. Bruker Application Note LCMS 186, www.bruker.com
We describe here a novel method for rapid Top Down analysis of N and O glycosylation in recombinant SARS CoV 2 RBDs utilizing Next Generation MALDI Top Down Sequencing (Next Gen MALDI TDS). Next Gen MALDI TDS combines MALDI In Source Decay (ISD) with ultrafast separation by Trapped Ion Mobility Spectrometry (TIMS), and high resolution, accurate time of flight mass analysis (TOF MS), delivering information simultaneously regarding primary sequence, terminal status and near terminal modifications of intact proteins.Summary: We describe here a novel method for rapid Top Down analysis of N and O glycosylation in recombinant SARS CoV 2 RBDs utilizing Next Generation MALDI Top Down Sequencing (Next Gen MALDI TDS).References: [1] C. Gstöttner, T. Zhang, A. Resemann, S. Ruben, S. Pengelley, D. Suckau, T. Welsink, M. Wuhrer, E. Domínguez Vega. Anal. Chem. 2021, 93, 17, 6839-6847
[2] A. Asperger, A. Resemann, W. Evers, N. Goedecke, D. Suckau. Bruker Application Note LCMS 186, www.bruker.com
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