An Integrated Protocol Based on Workflows of Imaged Capillary Isoelectric Focusing (Icief) for In-Depth Protein Heterogenous Characterization: High-Efficient Fractionation and Online Mass Spectrometry Detection DOI
Tao Bo,

Teresa Kwok,

Matt Countney

et al.

Published: Jan. 1, 2024

Language: Английский

Imaged capillary isoelectric focusing and online mass spectrometry for milk whey protein characterization in dairy products DOI

She Lin Chan,

Teresa Kwok,

Niusheng Xu

et al.

Analytical Biochemistry, Journal Year: 2025, Volume and Issue: 699, P. 115765 - 115765

Published: Jan. 6, 2025

Language: Английский

Citations

3
An integrated protocol based on workflows of imaged capillary isoelectric focusing (icIEF) for in-depth protein heterogenous characterization: high-efficient fractionation and online mass spectrometry detection DOI Creative Commons

Teresa Kwok,

She Lin Chan,

Mike Zhou

et al.

Green Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 100204 - 100204

Published: Jan. 1, 2025

Language: Английский

Citations

2
In-depth Characterization of a Cysteine-Linked ADC Disitamab Vedotin by Multiple LC-MS Analysis Methods and Cutting-Edge Imaged Capillary Isoelectric Focusing Coupled with Native Mass Spectrometry DOI
Gang Wu, Xiaoxi Zhang, Xin Wang

et al.

Journal of Chromatography A, Journal Year: 2024, Volume and Issue: 1736, P. 465353 - 465353

Published: Sept. 8, 2024

Language: Английский

Citations

5
Imaged Capillary Isoelectric Focusing Coupled to High‐Resolution Mass Spectrometry (icIEF‐MS) for Cysteine‐Linked Antibody–Drug Conjugate (ADC) Heterogeneity Characterization Under Native Condition DOI

Xiaoxi Zhang,

Gang Wu,

Min Du

et al.

Electrophoresis, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 30, 2024

ABSTRACT Native mass spectrometry (nMS) is a cutting‐edge technique that leverages electrospray ionization MS (ESI‐MS) to investigate large biomolecules and their complexes in solution. The goal of nMS retain the native structural features interactions analytes during transition gas phase, providing insights into natural conformations. In biopharmaceutical development, serves as powerful tool for analyzing complex protein heterogeneity, allowing examination non‐covalently bonded assemblies state closely resembles folded form. Herein, we present an imaged capillary isoelectric focusing–MS (icIEF–MS) workflow characterize cysteine‐linked antibody–drug conjugate (ADC) under conditions. Two ADCs were analyzed: latest generation ADC polatuzumab vedotin first FDA‐approved brentuximab vedotin. This benefits from recently developed icIEF system MS‐friendly capable directly coupling high‐sensitivity instrument. Results show separation influenced by both drug payloads post‐translational modifications (PTMs), which are then promptly identified MS. Overall, this icIEF–MS method demonstrates potential understand control critical quality attributes (CQAs) essential safe effective use ADCs.

Language: Английский

Citations

5
Evaluation of a cIEF Fractionation Workflow for Offline MS Analysis of Charge Variants of the Monoclonal Antibody Matuzumab DOI Creative Commons

Antonia Wittmann,

Yannick Wilke,

Nicolas Grammel

et al.

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

Published: Feb. 18, 2025

ABSTRACT Biological drugs like monoclonal antibodies require careful analysis and characterization to ensure product quality, safety, efficacy. Charge variants of the molecule are key interest analyzed using imaged capillary isoelectric focusing (icIEF). However, deeper these poses challenges. Two workflows for their exist: an ion‐exchange chromatography method variant collection before mass spectrometry (MS) analysis, which is labor‐intensive, direct coupling CE MS, allows detailed structural but has limitations, example, due incompatibilities with ES ionization high BGE concentrations. This study evaluates a platform that fractionates charge offline MS analysis. The suitability procedure in analytical icIEF methods converted into preparative cIEF fractionation by increasing sample concentration adding 20 mM arginine as cathodic spacer was tested. After chemical mobilization fraction collection, identity fractions determined fluorescence measurement reinjection protein‐containing fractions, previously developed method. subsequently performed. general workflow demonstrated Matuzumab. Transferring from 0.2 1.2 mg/mL yielded identical number peaks visually comparable peak profiles. separation took 50 min, additional 25 min 45 s per totaling approximately 2.5 h. Verification isolation straightforward via icIEF. Following fractionation, allowed identification main peaks. Preliminary results other indicated range experiments needs further investigation. Future work will aim optimize sensitivity, selectivity, time, reproducibility.

Language: Английский

Citations

0
Recent developments of imaged capillary isoelectric focusing technology for in-depth biopharmaceutical characterization DOI Creative Commons
Bo Tao, Feng Yang, Bo Yan

et al.

TrAC Trends in Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 118212 - 118212

Published: Feb. 1, 2025

Language: Английский

Citations

0
Advancing protein heterogeneity analysis: nano-flow pressure mobilization for precise icIEF fractionation and online MS detection DOI

Teresa Kwok,

She Lin Chan,

Niusheng Xu

et al.

Analytical Biochemistry, Journal Year: 2025, Volume and Issue: 701, P. 115825 - 115825

Published: March 2, 2025

Language: Английский

Citations

0
Advances and challenges in preparing membrane proteins for native mass spectrometry DOI

Cheng Di,

Yi Guo, Jixing Lyu

et al.

Biotechnology Advances, Journal Year: 2024, Volume and Issue: unknown, P. 108483 - 108483

Published: Nov. 1, 2024

Language: Английский

Citations

1
Retracted:An integrated protocol based on workflows of imaged capillary isoelectric focusing (icIEF) for in-depth protein heterogenous characterization: High-efficient fractionation and online mass spectrometry detection DOI Creative Commons

Teresa Kwok,

Matt Countney,

Tia Chan

et al.

Green Analytical Chemistry, Journal Year: 2024, Volume and Issue: 10, P. 100122 - 100122

Published: June 25, 2024

This article has been retracted: Please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/locate/withdrawalpolicy). retracted at the request of authors. The manuscript was submitted before a final round revision by As result, not all authors had opportunity to review and approve it, there are series errors that should have rectified prior publication. These are: • Some figures do contain reference original source (permissions obtained reproduce but this properly represented). sentences in Introduction were taken verbatim from other sources while no included source. need further supplement some data discussion improve paper. apologise for error.

Language: Английский

Citations

0
Imaged Capillary Isoelectric Focusing Coupled with Native Mass Spectrometry Combined with Other Separation Techniques for In-Depth Characterization of a Novel Cysteine-Linked Adc Disitamab Vedotin DOI
Gang Wu, Xiaoxi Zhang, Xin Wang

et al.

Published: Jan. 1, 2024

Language: Английский

Citations

0