Evaluation of a glycoengineered monoclonal antibody via LC-MS analysis in combination with multiple enzymatic digestion

MAbs
Renpeng LiuEllen P Guthrie

Abstract

Glycosylation affects the efficacy, safety and pharmacokinetics/pharmacodynamics properties of therapeutic monoclonal antibodies (mAbs), and glycoengineering is now being used to produce mAbs with improved efficacy. In this work, a glycoengineered version of rituximab was produced by chemoenzymatic modification to generate human-like N-glycosylation with α 2,6 linked sialic acid. This modified rituximab was comprehensively characterized by liquid chromatography-mass spectrometry and compared to commercially available rituximab. As anticipated, the majority of N-glycans were converted to α 2,6 linked sialic acid, in contrast to CHO-produced rituximab, which only contains α 2,3 linked sialic acid. Typical posttranslational modifications, such as pyro-glutamic acid formation at the N-terminus, oxidation at methionine, deamidation at asparagine, and disulfide linkages were also characterized in both the commercial and glycoengineered mAbs using multiple enzymatic digestion and mass spectrometric analysis. The comparative study reveals that the glycoengineering approach does not cause any additional posttranslational modifications in the antibody except the specific transformation of the glycoforms, demonstrating the mildness and ef...Continue Reading

References

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Citations

Oct 25, 2016·Trends in Biotechnology·Hye In ParkSang Taek Jung
Feb 1, 2018·International Journal of Molecular Sciences·Lindsay D BennettSylvain Marcel
May 29, 2018·Chemical Communications : Chem Comm·Sachin S ShivatareChi-Huey Wong
Oct 6, 2018·Glycobiology·Renato MastrangeliHorst Bierau
Mar 28, 2019·Annual Review of Biochemistry·Lai-Xi WangTiezheng Li
Jul 1, 2017·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Balazs BobalyDavy Guillarme

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Methods Mentioned

BETA
PCA
glycosylation
deamidation
transglycosylation

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