Advancements in Electrochemical Biosensors for Comprehensive Glycosylation Assessment of Biotherapeutics

Sensors, Journal Year: 2025, Volume and Issue: 25(7), P. 2064 - 2064

Published: March 26, 2025

Proteins represent a significant portion of the global therapeutics market, surpassing hundreds billions dollars annually. Among various post-translational modifications, glycosylation plays crucial role in influencing protein structure, stability, and function. This modification is especially important biotherapeutics, where precise characterization glycans vital for ensuring product efficacy safety. Although mass spectrometry-based techniques have become essential tools glycomic analysis due to their high sensitivity resolution, complexity lengthy processing times limit practical application. In contrast, electrochemical methods provide rapid, cost-effective, sensitive alternative assessment, enabling real-time glycan structures on biotherapeutic proteins. These techniques, often used conjunction with complementary methods, offer valuable insights into profiles both isolated glycoproteins intact cells. review examines latest advancements biosensors analysis, highlighting potential enhancing biotherapeutics advancing field precision medicine.

Language: Английский

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Standardized HILIC-FLD N-Glycan Analysis for Assessing N-Glycosylation Heterogeneity and Glycosylation-Related Quality Attributes in Bevacizumab from Multiple Manufacturers

Chromatographia, Journal Year: 2025, Volume and Issue: unknown

Published: April 11, 2025

Language: Английский

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N-linked glycosylation of the M-protein variable region: glycoproteogenomics reveals a new layer of personalized complexity in multiple myeloma

Clinical Chemistry and Laboratory Medicine (CCLM), Journal Year: 2024, Volume and Issue: 62(8), P. 1626 - 1635

Published: Feb. 9, 2024

Abstract Objectives Multiple myeloma (MM) is a plasma cell malignancy characterized by monoclonal expansion of cells that secrete characteristic M-protein. This M-protein crucial for diagnosis and monitoring MM in the blood patients. Recent evidence has emerged suggesting N-glycosylation variable (Fab) region contributes to pathogenicity, it risk factor disease progression disorders. Current methodologies lack specificity provide site-specific glycoprofile Fab regions M-proteins. Here, we introduce novel glycoproteogenomics method allows detailed glycoprofiling integrating patient specific sequences (genomics) with glycoproteomics. Methods Glycoproteogenomics was used analysis de novo sites First, Genomic identify sites. Subsequently glycopeptide LC-MS/MS glycan Results uncovered more than two-fold increase Light Chain M-proteins patients Myeloma compared polyclonal antibodies from healthy individuals. Subsequent 41 enrolled IFM 2009 clinical trial revealed majority were fully occupied complex type glycans, distinguishable Fc glycans due high levels sialylation, fucosylation bisecting structures. Conclusions Together, powerful tool study dyscrasias.

Language: Английский

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Effects of process intensification on homogeneity of an IgG1:κ monoclonal antibody during perfusion culture

Applied Microbiology and Biotechnology, Journal Year: 2024, Volume and Issue: 108(1)

Published: March 26, 2024

Abstract The pharmaceutical industry employs various strategies to improve cell productivity. These include process intensification, culture media improvement, clonal selection, supplementation and genetic engineering of cells. However, improved productivity has inherent risk impacting product quality attributes (PQA). PQAs may affect the products’ efficacy via stability, bioavailability, or in vivo bioactivity. Variations manufacturing introduce heterogeneity products by altering type extent N-glycosylation, which is a PQA therapeutic proteins. We investigated effect different densities representing increasing intensification perfusion on production an IgG1-κ monoclonal antibody from CHO-K1 line. This glycosylated both light chain heavy chain. Our results showed that contents glycosylation mAb increased G0F fucosylated glycans as group, whereas sialylated decreased, for whole protein. Overall, significant differences were observed amounts G0F, G1F, G0, G2FS1, G2FS2 across all levels. G2 N-glycans predominantly quantifiable rather than It be concluded there potential impact proteins during needs assessed. Since collected throughout duration process, lot allocation careful attention parameters, are affected critical parameters (CPPs). Key points • Molecular integrity suffer with intensity. Galactosylated decrease. Perfusion appears maintain protein charge structure.

Language: Английский

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Updates of the current strategies of labeling for N-glycan analysis

Journal of Chromatography B, Journal Year: 2024, Volume and Issue: 1237, P. 124068 - 124068

Published: Feb. 27, 2024

Language: Английский

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Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

Mass Spectrometry Reviews, Journal Year: 2024, Volume and Issue: unknown

Published: June 24, 2024

Abstract The use of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry for the analysis carbohydrates and glycoconjugates is a well‐established technique this review 12 th update original article published in 1999 brings coverage literature to end 2022. As with previous review, also includes few papers that describe methods appropriate by MALDI, such as sample preparation, even though ionization method not MALDI. follows same format reviews. It divided into three sections: (1) general aspects theory MALDI process, matrices, derivatization, imaging, fragmentation, quantification computer software structural identification. (2) Applications various types oligo‐ polysaccharides, glycoproteins, glycolipids, glycosides biopharmaceuticals, (3) other areas medicine, industrial processes, natural products glycan synthesis where extensively used. Much material relating applications presented tabular form. still an ideal carbohydrate analysis, particularly its ability produce single ions from each analyte advancements range show little sign diminishing.

Language: Английский

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Purification free N-glycan analysis by capillary zone electrophoresis: Hunt for the lost glycans

Journal of Pharmaceutical and Biomedical Analysis, Journal Year: 2023, Volume and Issue: 238, P. 115812 - 115812

Published: Oct. 21, 2023

Capillary gel electrophoresis is a widely used method for rapid separation of fluorophore labeled carbohydrates. Even though, many publications conferred about this popular technique, no report yet investigated the possible sample losses during purification process labeling reaction mixture. In present work, normal polarity capillary zone mode was applied to take advantage opposite migration directions electroosmotic flow and negatively charged components using Tris-hexanoic acid running buffer at basic pH. For free oligosaccharide analysis, parameters were designed in such way that triple reagent aminopyrenetrisulfonate (APTS) could not enter contrary interest, therefore, APTS did have be removed before analysis. The show electrophoretic profile differences possibly caused by cleanup immediately apparent comparing electropherograms purified non-purified maltooligosaccharides. Furthermore, qualitative quantitative N-glycosylation alterations revealed CZE mixtures after along with analysis consecutively washing solutions well characterized standard glycoproteins IgG, ribonuclease B fetuin.

Language: Английский

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Relative quantitation of glycans in cetuximab using ultra-high-performance liquid chromatography-high-resolution mass spectrometry by Pronase E digestion

Journal of Chromatography A, Journal Year: 2022, Volume and Issue: 1677, P. 463302 - 463302

Published: July 5, 2022

Language: Английский

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N-glycosylation reinforces interaction of immune checkpoint TIM-3 with a small molecule ligand

Computational Biology and Chemistry, Journal Year: 2023, Volume and Issue: 104, P. 107852 - 107852

Published: March 22, 2023

Language: Английский

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Recent progress in high-throughput and automated characterization of N-glycans in monoclonal antibodies

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 169, P. 117397 - 117397

Published: Oct. 29, 2023

Language: Английский

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