Acceptable changes in quality attributes of glycosylated biopharmaceuticals

Schiestl, Martin; Stangler, Thomas; Torella, Claudia; Čepeljnik, Tadej; Toll, Hansjörg; Grau, Roger

doi:10.1038/nbt.1839

Cited by 441 publications

(351 citation statements)

References 9 publications

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“…It only represents 2-5% of the total mass of the protein but it is subjected to extensive studies due to its significant influence on effector functions of mAbs, especially antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) [5][6][7][8]. As a consequence, the mAbs glycosylation profile is considered as a critical quality attribute (CQA) and must be thoroughly analyzed [9][10][11][12][13]. The complexity and heterogeneity of the glycosylation pattern is mainly due to mAbs production in living expression systems [14][15][16] and requires a number of orthogonal analytical techniques to be fully characterized.…”

Section: Introductionmentioning

confidence: 99%

Monoclonal antibody N-glycosylation profiling using capillary electrophoresis – Mass spectrometry: Assessment and method validation

Giorgetti

D’Atri

Canonge

et al. 2018

Talanta

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A B S T R A C TCharacterization of therapeutic proteins represents a major challenge for analytical sciences due to their heterogeneity caused by post-translational modifications (PTM). Among these PTM, glycosylation which is possibly the most prominent, require comprehensive identification because of their major influence on protein structure and effector functions of monoclonal antibodies (mAbs). As a consequence, glycosylation profiling must be deeply characterized. For this application, several analytical methods such as separation-based or MS-based methods, were evaluated. However, no CE-ESI-MS approach has been assessed and validated. Here, we illustrate how the use of CE-ESI-MS method permits the comprehensive characterization of mAbs N-glycosylation at the glycopeptide level to perform relative quantitation of N-glycan species. Validation of the CE-ESI-MS method in terms of robustness and reproducibility was demonstrated through the relative quantitation of glycosylation profiles for ten different mAbs produced in different cell lines. Glycosylation patterns obtained for each mAbs were compared to Hydrophilic Interaction Chromatography of 2-aminobenzamide labelled glycans with fluorescence detector (HILIC-FD) analysis considered as a reference method. Very similar glycoprofiling were obtained with the CE-ESI-MS and HILIC-FD demonstrating the attractiveness of CE-ESI-MS method to characterize and quantify the glycosylation heterogeneity of a wide range of therapeutic mAbs with high accuracy and precision.

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Section: Introductionmentioning

confidence: 99%

Monoclonal antibody N-glycosylation profiling using capillary electrophoresis – Mass spectrometry: Assessment and method validation

Giorgetti

D’Atri

Canonge

et al. 2018

Talanta

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“…Indeed, substantial differences in pharmacokinetics and biological activities have been reported for products undergoing changes in manufacturing processes (24). However, variations in extravasation rates and disease-homing properties may often go unnoticed, unless quantitative biodistribution studies are performed.…”

Section: Resultsmentioning

confidence: 99%

Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies

Venetz¹,

Hess²,

Lin

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The ability of antibodies to extravasate out of blood vessels is critical for therapeutic activity, because molecular targets for most diseases are located outside of the endothelial lining. By performing detailed biodistribution studies with a novel IL9-armed cancer-specific antibody, we identified a clear correlation between N-linked glycan structures and tumor-targeting efficiencies. Site-specific glycan analysis provided a detailed view of the glycan microheterogeneity present on the IL9 portion of the recombinant protein. Nonsialylated glycan structures have a negative impact on disease-homing activity, highlighting the importance of glycosylation control and characterization during process development.

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“…On the other hand, N-terminal Gln cyclization to pyroglutamate results in more acidic antibodies with lower molecular weight following the removal of NH 3 159 . There is no evidence that both C-terminal Lys and N-terminal Gln processing impact the biological activity of the molecule 159,160 or affect immunogenicity or safety 159 . Furthermore, C-terminal proline (Pro) amidation of monoclonal antibodies has been described 44,161,162 .…”

Section: C-and N-terminal Modificationsmentioning

confidence: 99%

Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

Biotechnology Progress

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Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

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Acceptable changes in quality attributes of glycosylated biopharmaceuticals

Cited by 441 publications

References 9 publications

Monoclonal antibody N-glycosylation profiling using capillary electrophoresis – Mass spectrometry: Assessment and method validation

Monoclonal antibody N-glycosylation profiling using capillary electrophoresis – Mass spectrometry: Assessment and method validation

Glycosylation profiles determine extravasation and disease-targeting properties of armed antibodies

Tailoring recombinant protein quality by rational media design

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