“Small is beautiful”—Examining reliable determination of low-abundant therapeutic antibody glycovariants

Katharina B&#246;ttinger; Christof Regl; Veronika Sch&#228;pert&#246;ns; Erdmann Rapp; Therese Wohlschlager; Christian G. Huber

doi:10.1016/j.jpha.2024.100982

Volume 14 Issue 10

Oct. 2024

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Katharina Böttinger, Christof Regl, Veronika Schäpertöns, Erdmann Rapp, Therese Wohlschlager, Christian G. Huber. “Small is beautiful”—Examining reliable determination of low-abundant therapeutic antibody glycovariants[J]. Journal of Pharmaceutical Analysis, 2024, 14(10): 100982. doi: 10.1016/j.jpha.2024.100982

Citation:

Katharina Böttinger, Christof Regl, Veronika Schäpertöns, Erdmann Rapp, Therese Wohlschlager, Christian G. Huber. “Small is beautiful”—Examining reliable determination of low-abundant therapeutic antibody glycovariants[J]. Journal of Pharmaceutical Analysis, 2024, 14(10): 100982. doi: 10.1016/j.jpha.2024.100982

Citation:

Katharina Böttinger, Christof Regl, Veronika Schäpertöns, Erdmann Rapp, Therese Wohlschlager, Christian G. Huber. “Small is beautiful”—Examining reliable determination of low-abundant therapeutic antibody glycovariants[J]. Journal of Pharmaceutical Analysis, 2024, 14(10): 100982. doi: 10.1016/j.jpha.2024.100982

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“Small is beautiful”—Examining reliable determination of low-abundant therapeutic antibody glycovariants

doi: 10.1016/j.jpha.2024.100982

a. Department of Biosciences and Medical Biology, Bioanalytical Research Labs, University of Salzburg, Salzburg, 5020, Austria;
b. Center for Tumorbiology and Immunology (CTBI), University of Salzburg, Salzburg, 5020, Austria;
c. glyXera GmbH, Magdeburg, Sachsen-Anhalt, 39014, Germany;
d. Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, 39106, Germany

Funds:

This research was funded in whole or in part by the Austrian Science Fund (FWF) (FG 1201 N, "DigiTherapeutX"],. Figures were created in Inkscape (v1.3.1). Fig.

Received Date: Jan. 09, 2024
Accepted Date: Apr. 23, 2024
Rev Recd Date: Mar. 21, 2024
Publish Date: Apr. 26, 2024

Abstract

Abstract

Glycans associated with biopharmaceutical drugs play crucial roles in drug safety and efficacy, and therefore, their reliable detection and quantification is essential. Our study introduces a multi-level quantification approach for glycosylation analysis in monoclonal antibodies (mAbs), focusing on minor abundant glycovariants. Mass spectrometric data is evaluated mainly employing open-source software tools. Released N-glycan and glycopeptide data form the basis for integrating information across different structural levels up to intact glycoproteins. Comprehensive comparison showed that indeed, variations across structural levels were observed especially for minor abundant species. Utilizing modification finder (MoFi), a tool for annotating mass spectra of intact proteins, we quantify isobaric glycosylation variants at the intact protein level. Our workflow's utility is demonstrated on NISTmAb, rituximab and adalimumab, profiling their minor abundant variants for the first time across diverse structural levels. This study enhances understanding and accessibility in glycosylation analysis, spotlighting minor abundant glycovariants in therapeutic antibodies.
- Glycosylation,
- Mass spectrometry,
- Monoclonal antibodies,
- Abundance profiling,
- Minor glycovariants,
- Multi-level analysis,
- Data integration

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References(66)

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