Evaluation of oligosaccharide methods for carbohydrate analysis in a fully human monoclonal antibody and comparison of the results to the monosaccharide composition determination by a novel calculation

Adamo, Michael; Qiu, Difei; Dick, Lawrence W.; Zeng, Mingyong; Lee, An-Horng; Cheng, Kuang-Chuan

doi:10.1016/j.jpba.2008.10.008

Cited by 20 publications

(10 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

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

“…The addition of orthogonal methods provides complementary and corroboratory information to clarify instances of inconclusive data. High-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and hydrophilic-interaction chromatography with fluorescence detection (HILIC-FD) are among the most widely used and informative methods for the determination of glycoprotein glycosylation, and are industry standards in glycan analysis [53,[55][56][57][58][59][60][61][62][63][64]. However, both HPAEC-PAD and HILIC-FD require removal of glycans from the underlying protein prior to analysis, involve multiple steps are overall rather time-consuming and relatively expensive, demand significant operator experience and expertise, and involve considerable data analysis.…”

Section: Introductionmentioning

confidence: 99%

Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody

Cook

Kaldas

Muradia

et al. 2015

Journal of Chromatography B

View full text Add to dashboard Cite

“…The first investigates glycopeptides, which requires the protein backbone to be cleaved-usually with trypsin to yield low molecular weight glycopeptides, which are then analyzed by the techniques described later in this section. 32,119,120,147 The second strategy is to remove the oligosaccharides from the protein backbone. This can be achieved by chemical means (e.g., hydrazinolysis); however, enzymatic cleavage is more frequently used.…”

Section: Techniques For Mab Glycosylation Analysismentioning

confidence: 99%

“…This combination of analytical techniques permits separation and absolute quantification of oligosaccharide species by LC or CE (through fluorescence or absorption) and identification (or identity confirmation) of each species which has been separated by mass spectrometry. Examples of the combination of these techniques towards the characterization of mAb glycans are: CE-LIF-ESI-MS, 164,166 RP-HPLC-ESI-MS. 49,82,[117][118][119][120]147,150,153,155,164,167 …”

Section: Combination Of Analytical Techniques (Lc-ms and Ce-ms)mentioning

confidence: 99%

Towards the implementation of quality by design to the production of therapeutic monoclonal antibodies with desired glycosylation patterns

Val

Kontoravdi

Nagy

2010

Biotechnology Progress

126

View full text Add to dashboard Cite

Quality by design (QbD) is a scheme for the development, manufacture, and approval of pharmaceutical products. The end goal of QbD is to ensure product quality by building it into the manufacturing process. The main regulatory bodies are encouraging its implementation to the manufacture of all new pharmaceuticals including biological products. Monoclonal antibodies (mAbs) are currently the leading products of the biopharmaceutical industry. It has been widely reported that glycosylation directly influences the therapeutic mechanisms by which mAbs function in vivo. In addition, glycosylation has been identified as one of the main sources of monoclonal antibody heterogeneity, and thus, a critical parameter to follow during mAb manufacture. This article reviews the research on glycosylation of mAbs over the past 2 decades under the QbD scope. The categories presented under this scope are: (a) definition of the desired clinical effects of mAbs, (b) definition of the glycosylation-associated critical quality attributes (glycCQAs) of mAbs, (c) assessment of process parameters that pose a risk for mAb glycCQAs, and (d) methods for accurately quantifying glycCQAs of mAbs. The information available in all four areas leads us to conclude that implementation of QbD to the manufacture of mAbs with specific glycosylation patterns will be a reality in the near future. We also foresee that the implementation of QbD will lead to the development of more robust and efficient manufacturing processes and to a new generation of mAbs with increased clinical efficacy.

show abstract

Evaluation of oligosaccharide methods for carbohydrate analysis in a fully human monoclonal antibody and comparison of the results to the monosaccharide composition determination by a novel calculation

Cited by 20 publications

References 24 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

Comparison of orthogonal chromatographic and lectin-affinity microarray methods for glycan profiling of a therapeutic monoclonal antibody

Towards the implementation of quality by design to the production of therapeutic monoclonal antibodies with desired glycosylation patterns

Contact Info

Product

Resources

About