Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis

The identification and accurate quantitation of the various glycoforms contained in therapeutic monoclonal antibodies (mAbs) is one of the main analytical needs in the biopharmaceutical industry, and glycosylation represents a crucial critical quality attribute (CQA) that needs to be addressed. Curr...

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Autores principales:	Bastiaan L. Duivelshof, Steffy Denorme, Koen Sandra, Xiaoxiao Liu, Alain Beck, Matthew A. Lauber, Davy Guillarme, Valentina D’Atri
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	glycan profiling quantitative analysis HILIC-MS protein subunits monoclonal antibodies Pharmacy and materia medica RS1-441
Acceso en línea:	https://doaj.org/article/9ef3402fc0064b6fa7968bd0b3ad2485
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spelling	oai:doaj.org-article:9ef3402fc0064b6fa7968bd0b3ad24852021-11-25T18:40:19ZQuantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis10.3390/pharmaceutics131117441999-4923https://doaj.org/article/9ef3402fc0064b6fa7968bd0b3ad24852021-10-01T00:00:00Zhttps://www.mdpi.com/1999-4923/13/11/1744https://doaj.org/toc/1999-4923The identification and accurate quantitation of the various glycoforms contained in therapeutic monoclonal antibodies (mAbs) is one of the main analytical needs in the biopharmaceutical industry, and glycosylation represents a crucial critical quality attribute (CQA) that needs to be addressed. Currently, the reference method for performing such identification/quantitation consists of the release of the <i>N</i>-glycan moieties from the mAb, their labelling with a specific dye (e.g., 2-AB or RFMS) and their analysis by HILIC-FLD or HILIC-MS. In this contribution, the potential of a new cost- and time-effective analytical approach performed at the protein subunit level (middle-up) was investigated for quantitative purposes and compared with the reference methods. The robustness of the approach was first demonstrated by performing the relative quantification of the glycoforms related to a well characterized mAb, namely adalimumab. Then, the workflow was applied to various glyco-engineered mAb products (i.e., obinutuzumab, benralizumab and atezolizumab). Finally, the glycosylation pattern of infliximab (Remicade<sup>®</sup>) was assessed and compared to two of its commercially available biosimilars (Remsima<sup>®</sup> and Inflectra<sup>®</sup>). The middle-up analysis proved to provide accurate quantitation results and has the added potential to be used as multi-attribute monitoring method.Bastiaan L. DuivelshofSteffy DenormeKoen SandraXiaoxiao LiuAlain BeckMatthew A. LauberDavy GuillarmeValentina D’AtriMDPI AGarticleglycan profilingquantitative analysisHILIC-MSprotein subunitsmonoclonal antibodiesPharmacy and materia medicaRS1-441ENPharmaceutics, Vol 13, Iss 1744, p 1744 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	glycan profiling quantitative analysis HILIC-MS protein subunits monoclonal antibodies Pharmacy and materia medica RS1-441
spellingShingle	glycan profiling quantitative analysis HILIC-MS protein subunits monoclonal antibodies Pharmacy and materia medica RS1-441 Bastiaan L. Duivelshof Steffy Denorme Koen Sandra Xiaoxiao Liu Alain Beck Matthew A. Lauber Davy Guillarme Valentina D’Atri Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis
description	The identification and accurate quantitation of the various glycoforms contained in therapeutic monoclonal antibodies (mAbs) is one of the main analytical needs in the biopharmaceutical industry, and glycosylation represents a crucial critical quality attribute (CQA) that needs to be addressed. Currently, the reference method for performing such identification/quantitation consists of the release of the <i>N</i>-glycan moieties from the mAb, their labelling with a specific dye (e.g., 2-AB or RFMS) and their analysis by HILIC-FLD or HILIC-MS. In this contribution, the potential of a new cost- and time-effective analytical approach performed at the protein subunit level (middle-up) was investigated for quantitative purposes and compared with the reference methods. The robustness of the approach was first demonstrated by performing the relative quantification of the glycoforms related to a well characterized mAb, namely adalimumab. Then, the workflow was applied to various glyco-engineered mAb products (i.e., obinutuzumab, benralizumab and atezolizumab). Finally, the glycosylation pattern of infliximab (Remicade<sup>®</sup>) was assessed and compared to two of its commercially available biosimilars (Remsima<sup>®</sup> and Inflectra<sup>®</sup>). The middle-up analysis proved to provide accurate quantitation results and has the added potential to be used as multi-attribute monitoring method.
format	article
author	Bastiaan L. Duivelshof Steffy Denorme Koen Sandra Xiaoxiao Liu Alain Beck Matthew A. Lauber Davy Guillarme Valentina D’Atri
author_facet	Bastiaan L. Duivelshof Steffy Denorme Koen Sandra Xiaoxiao Liu Alain Beck Matthew A. Lauber Davy Guillarme Valentina D’Atri
author_sort	Bastiaan L. Duivelshof
title	Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis
title_short	Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis
title_full	Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis
title_fullStr	Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis
title_full_unstemmed	Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis
title_sort	quantitative <i>n-</i>glycan profiling of therapeutic monoclonal antibodies performed by middle-up level hilic-hrms analysis
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/9ef3402fc0064b6fa7968bd0b3ad2485
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Quantitative <i>N-</i>Glycan Profiling of Therapeutic Monoclonal Antibodies Performed by Middle-Up Level HILIC-HRMS Analysis

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