High-Throughput Glycomic Methods

Chemical Reviews, Journal Year: 2022, Volume and Issue: 122(20), P. 15865 - 15913

Published: July 7, 2022

Glycomics aims to identify the structure and function of glycome, complete set oligosaccharides (glycans), produced in a given cell or organism, as well genes other factors that govern glycosylation. This challenging endeavor requires highly robust, sensitive, potentially automatable analytical technologies for analysis hundreds thousands glycomes timely manner (termed high-throughput glycomics). review provides historic overview highlights recent developments challenges glycomic profiling by most prominent approaches, with N-glycosylation focal point. It describes current state-of-the-art regarding levels characterization widely used technologies, selected applications glycomics deciphering glycosylation process healthy disease states, future perspectives.

Language: Английский

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Capillary electrophoresis and the biopharmaceutical industry: Therapeutic protein analysis and characterization

TrAC Trends in Analytical Chemistry, Journal Year: 2021, Volume and Issue: 144, P. 116407 - 116407

Published: Aug. 25, 2021

Capillary electrophoresis (CE) has emerged as a powerful technique for comprehensive physicochemical characterization of monoclonal antibodies (mAbs) well other therapeutic modalities. The method provides high resolution separation and sensitivity analysis biomolecules. CE based techniques such sodium dodecyl sulphate capillary gel (SDS-CGE, also referred to SDS), zone (CZE) imaged isoelectric focusing (iCIEF) have been increasingly adopted assess size heterogeneity, glycosylation heterogeneity charge in mAbs related This paper reviews the latest application developments methods routine release testing, stability testing in-depth characterization. In addition, advantages disadvantages each these are critically discussed.

Language: Английский

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Applications of capillary electromigration methods for separation and analysis of proteins (2017–mid 2021) – A review

Analytica Chimica Acta, Journal Year: 2022, Volume and Issue: 1209, P. 339447 - 339447

Published: Jan. 7, 2022

Language: Английский

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Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications—Updated and completely revised edition

Electrophoresis, Journal Year: 2023, Volume and Issue: 44(17-18), P. 1279 - 1341

Published: Aug. 3, 2023

Abstract This review is in support of the development selective, precise, fast, and validated capillary electrophoresis (CE) methods. It follows up a similar article from 1998, Wätzig H, Degenhardt M, Kunkel A. “Strategies for electrophoresis: method validation pharmaceutical biological applications,” pointing out which fundamentals are still valid at same time showing enormous achievements last 25 years. The structures both reviews widely similar, order to facilitate their simultaneous use. Focusing on applications, successful use CE now demonstrated by more than 600 carefully selected references. Many those recent reviews; therefore, significant overview about field provided. There extra sections sample pretreatment related microchip CE, completely revised section protein analytes biomolecules general. general strategies summed with regard selectivity, efficiency, precision, analysis time, limit detection, requirements, validation.

Language: Английский

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Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

Journal of Separation Science, Journal Year: 2023, Volume and Issue: 46(15)

Published: May 26, 2023

Capillary electrophoresis is recognized as a valued separation technique for its high efficiency, low sample consumption, good economic and ecological aspects, reproducibility, complementarity to traditional liquid chromatography techniques. experiments are generally performed utilizing optical detection, such ultraviolet or fluorescence detectors. However, in order provide structural information, capillary hyphenated highly sensitive selective mass spectrometry has been developed overcome the limitations of detections. electrophoresis‐mass increasingly popular protein analysis, including biopharmaceutical biomedical research. It frequently applied determination physicochemical biochemical parameters proteins, offers excellent performance in‐depth characterizations biopharmaceuticals at various levels also already proven promising tool biomarker discovery. In this review, we focus on possibilities analysis their intact level. Various modes interfaces, well approaches prevent adsorption enhance loading capacity, discussed recent (2018–March 2023) developments applications field summarized.

Language: Английский

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Current green capillary electrophoresis and liquid chromatography methods for analysis of pharmaceutical and biomedical samples (2019–2023) – A review

Analytica Chimica Acta, Journal Year: 2024, Volume and Issue: 1323, P. 342889 - 342889

Published: June 20, 2024

Separation analytical methods, including liquid chromatography (LC) and capillary electrophoresis (CE), in combination with an appropriate detection technique, are dominant powerful approaches preferred the analysis of pharmaceutical biomedical samples. Recent trends methods focused on activities that push them to field greenness sustainability. New based implementation greener solvents, non-hazardous chemicals, reagents have grown exponentially. Similarly, recent pushed strategies miniaturization, reduction wastes, avoiding derivatization procedures, or energy consumption. However, real method can be evaluated only according objective sufficient metric offering complex results taking into account all twelve rules green chemistry (SIGNIFICANCE mnemonic system). This review provides extensive overview papers published area development LC CE over last 5 years (2019-2023). The main focus is situated metrics used for evaluation applied determination bioactive agents. It critically evaluates compares demands applicability quality control clinical environment requirements (GAC). Greenness practicality summarized re-evaluated newly use tools, i.e., Analytical GREEnness (AGREE), Green Procedure Index (GAPI), Blue Applicability Grade (BAGI), Sample Preparation Metric Sustainability (SPMS). Moreover, general conclusions future perspectives greening procedures systems presented. paper should provide comprehensive information chemists, biochemists, it also represent a valuable source clinicians, laboratories interested greenness, practicality,

Language: Английский

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Multi‐dimensional technology – Recent advances and applications for biotherapeutic characterization

Journal of Separation Science, Journal Year: 2024, Volume and Issue: 47(5)

Published: March 1, 2024

This review provides an overview of the latest advancements and applications in multi‐dimensional liquid chromatography coupled with mass spectrometry (mD‐LC‐MS), covering aspects such as inter‐laboratory studies, digestion strategy, trapping column, multi‐level analysis. The shift from offline to online workflow reduces sample processing artifacts, analytical variability, analysis time, labor required for data acquisition. Over past few years, this technique has demonstrated sufficient maturity application across a diverse range complex products. Moreover, there is potential strategy evolve into integrated process technology tool real‐time monitoring monoclonal antibody quality. also identifies emerging trends, including its new modalities, possibility evaluating biological activity within mD‐LC set‐up, consideration capillary electrophoresis alternative mD‐LC. As mD‐LC‐MS continues integrate it holds shape next generation tools, offering exciting possibilities enhanced characterization biopharmaceutical

Language: Английский

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Applications of capillary electrophoresis for biopharmaceutical product characterization

Electrophoresis, Journal Year: 2021, Volume and Issue: 43(1-2), P. 143 - 166

Published: Sept. 30, 2021

Capillary electrophoresis (CE), after being introduced several decades ago, has carved out a niche for itself in the field of analytical characterization biopharmaceutical products. It does not only offer fast separation, high resolution miniaturized format, but equally importantly represents an orthogonal separation mechanism to high-performance liquid chromatography. Therefore, it is surprising that CE-based methods can be found all major pharmacopoeias and are recommended analysis products during process development, characterization, quality control, release testing. Different formats CE, such as capillary gel electrophoresis, isoelectric focusing, zone widely used size charge heterogeneity well purity stability testing therapeutic proteins. Hyphenation CE with MS emerging promising bioanalytical tool assess primary structure proteins along any impurities. In this review, we confer latest developments critical attributes covering past 6 years (2015-2021). Monoclonal antibodies, due their significant share market, have been given prioritized coverage.

Language: Английский

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Two-Dimensional Capillary Zone Electrophoresis-Mass Spectrometry: Intact mAb Charge Variant Separation Followed by Peptide Level Analysis Using In-Capillary Digestion

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(8), P. 4059 - 4066

Published: Feb. 17, 2023

Characterization of charge heterogeneity is an essential pillar for pharmaceutical development and quality control therapeutic monoclonal antibodies (mAbs). The highly selective commonly applied capillary zone electrophoresis (CZE) method containing high amounts ε-aminocaproic acid (EACA) provides a detailed robust profile intact mAb variants. Nevertheless, the exact location protein modifications within these profiles remains ambiguous. Electrospray ionization mass spectrometry (ESI-MS) promising tool this purpose; however, EACA incompatible with electrospray. In context, we present two-dimensional CZE-CZE-MS system to combine efficient variant separation mAbs subsequent peptide analysis after in-capillary digestion selected first dimension based on generic CZE(EACA) in fused silica capillary. second dimension, neutral-coated used reduction Tris(2-carboxyethyl)phosphine (TCEP) pepsin, followed by CZE MS/MS-characterization resulting peptides. setup demonstrated using stressed nonstressed where peaks basic, main, acidic variants were transferred heart-cut fashion, digested, characterized level. Sequence coverages more than 90% obtained heavy chain (HC) light (LC) four different mAbs, including low-abundance (<2% main peak). Frequently observed (deamidation, oxidation, etc.) could be detected localized. This study demonstrates proof-of-concept identification localization from and, way, expected support testing pharmaceuticals.

Language: Английский

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Recent capillary electrophoresis applications for upstream and downstream biopharmaceutical process monitoring

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 160, P. 116975 - 116975

Published: Feb. 10, 2023

The biopharmaceutical market is one of the fastest growing biotechnology markets. In order to ensure affordable and reliable therapeutics, process has be closely monitored. Capillary electrophoresis (CE) proven a valuable technique for analysis product concentration, critical quality attributes, process-related impurities, nutrients metabolites in cell culture medium. zone electrophoresis, capillary gel isoelectric focusing are extensively used size or charge heterogeneity determination. CE number benefits upstream downstream process-intermediates, including ability handle highly complex matrices found high resolving power, little sample preparation requirements, rapid analysis, low solvent consumption. small volumes (nL range) especially beneficial microbioreactor clone selection experiments. simple setup possibility miniaturisation automation using microchip provides great opportunities on-site, real-time monitoring process. This review discusses applications processing last decade.

Language: Английский

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