Glycan analysis of therapeutic glycoproteins

Zhang, Lei; Luo, Shen; Zhang, Baolin

doi:10.1080/19420862.2015.1117719

Cited by 154 publications

(129 citation statements)

References 114 publications

Supporting

Mentioning

129

Contrasting

Order By: Relevance

“…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. Several analytical methods have been described for the glyco-variants characterization at different levels (from released glycans to intact protein level) including separative techniques (liquid chromatography (LC), capillary electrophoresis (CE)) often coupled to spectrometric, amperometric and mass spectrometric detection [17][18][19][20][21]. Recently, Reusch's group published two major articles dealing with the analysis of Fc-glycosylation profiles, and comparing several separation methods hyphenated or not with mass spectrometry (MS) detection [20,21].…”

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

View full text Add to dashboard Cite

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.

show abstract

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

View full text Add to dashboard Cite

show abstract

“…This enabled the novel identification of 70 glycans and 4 never before detected glycoforms. Other aspects of product quality can be determined with unprecedented detail, including post‐translational modifications, glycan content, and structure …”

Section: Proteomicsmentioning

confidence: 99%

CHO‐Omics Review: The Impact of Current and Emerging Technologies on Chinese Hamster Ovary Based Bioproduction

Stolfa

Smonskey

Boniface

et al. 2017

Biotechnology Journal

View full text Add to dashboard Cite

CHO cells are the most prevalent platform for modern bio-therapeutic production. Currently, there are several CHO cell lines used in bioproduction with distinct characteristics and unique genotypes and phenotypes. These differences limit advances in productivity and quality that can be achieved by the most common approaches to bioprocess optimization and cell line engineering. Incorporating omics-based approaches into current bioproduction processes will complement traditional methodologies to maximize gains from CHO engineering and bioprocess improvements. In order to highlight the utility of omics technologies in CHO bioproduction, the authors discuss current applications as well as limitations of genomics, transcriptomics, proteomics, metabolomics, lipidomics, fluxomics, glycomics, and multi-omics approaches and the potential they hold for the future of bioproduction. Multiple omics approaches are currently being used to improve CHO bioprocesses; however, the application of these technologies is still limited. As more CHO-omic datasets become available and integrated into systems models, the authors expect significant gains in product yield and quality. While individual omics technologies provide incremental improvements in bioproduction, the authors will likely see the most significant gains by applying multi-omics and systems biology approaches to individual CHO cell lines.

show abstract

“…Although MS‐based methods can reliably identify the structure, linkage, and position of glycans, enzymatic or chemical stripping of glycans from proteins prior to MS profiling prevents accurate detection and identification of total glycans. Moreover, these methods are usually time‐consuming and require complex sample preparation procedures …”

Section: Introductionmentioning

confidence: 99%

“…Lectins are a group of carbohydrate‐binding proteins that specifically bind different glycans. The advantages of using lectin microarray over traditional MS‐based methods include the simplicity and high sensitivity of the method that supports direct global glycomic profiling, the lower stringency of initial sample purity (crude glycoprotein samples can be analyzed), and the comparatively simple sample preparation procedure (without glycan‐release and purification) . As protein fragmentation or glycan liberation is not required during sample preparation, the sampled glycoproteins can retain their intact natural conformations and abundance.…”

Section: Introductionmentioning

confidence: 99%

Application of Lectin Microarrays for Biomarker Discovery

et al. 2020

View full text Add to dashboard Cite

Many proteins in living organisms are glycosylated. As their glycan patterns exhibit protein‐, cell‐, and tissue‐specific heterogeneity, changes in the glycosylation levels could serve as useful indicators of various pathological and physiological states. Thus, the identification of glycoprotein biomarkers from specific changes in the glycan profiles of glycoproteins is a trending field. Lectin microarrays provide a new glycan analysis platform, which enables rapid and sensitive analysis of complex glycans without requiring the release of glycans from the protein. Recent developments in lectin microarray technology enable high‐throughput analysis of glycans in complex biological samples. In this review, we will discuss the basic concepts and recent progress in lectin microarray technology, the application of lectin microarrays in biomarker discovery, and the challenges and future development of this technology. Given the tremendous technical advancements that have been made, lectin microarrays will become an indispensable tool for the discovery of glycoprotein biomarkers.

show abstract

Glycan analysis of therapeutic glycoproteins

Cited by 154 publications

References 114 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

CHO‐Omics Review: The Impact of Current and Emerging Technologies on Chinese Hamster Ovary Based Bioproduction

Application of Lectin Microarrays for Biomarker Discovery

Contact Info

Product

Resources

About