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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Capillary electrophoresis‐mass spectrometry for protein analyses under native conditions: Current progress and perspectives

PROTEOMICS, Journal Year: 2023, Volume and Issue: 24(3-4)

Published: June 13, 2023

Abstract Native mass spectrometry is a rapidly emerging technique for fast and sensitive structural analysis of protein constructs, maintaining the higher order structure. The coupling with electromigration separation techniques under native conditions enables characterization proteoforms highly complex mixtures. In this review, we present an overview current CE‐MS technology. First, status described capillary zone electrophoresis (CZE), affinity (ACE), isoelectric focusing (CIEF), as well their chip‐based formats, including essential parameters such electrolyte composition coatings. Further, required ESI‐MS (large) instrumental QTOF Orbitrap systems, requirements interfacing are presented. On basis, methods applications different modes summarized discussed in context biological, medical, biopharmaceutical questions. Finally, key achievements highlighted concluded, while remaining challenges pointed out.

Language: Английский

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Performance of capillary electrophoresis instruments – State of the art and outlook

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 163, P. 117056 - 117056

Published: April 13, 2023

Language: Английский

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Transient isotachophoresis–Capillary zone electrophoresis–Mass spectrometry method with off-line microscale solid phase extraction pretreatment for quantitation of intact low molecular mass proteins in various biological fluids

Journal of Chromatography A, Journal Year: 2024, Volume and Issue: 1718, P. 464697 - 464697

Published: Feb. 3, 2024

Quantification of proteins is still predominantly done by the traditional bottom-up approach. Targeting intact in complex biological matrices connected with multiple challenges during sample pretreatment, separation, and detection step analytical workflow. In this work, we focused on development an on-line hyphenated capillary zone electrophoresis-mass spectrometry method employing off-line microscale solid-phase extraction based hydrophilic lipophilic balance (HLB) sorbent as a pretreatment for analysis low molecular mass (<20 kDa) spiked various fluids (human serum, plasma, urine, saliva). A detailed optimization process involved selection suitable surface, background electrolyte (BGE), comparison two in-capillary preconcentration methods, namely transient isotachophoresis (tITP) dynamic pH junction (DPJ), to enhance sensitivity method. Optimum separation analytes was achieved using uncoated bare fused silica 500 mM formic acid (pH 1.96) + 5% (v/v) acetonitrile BGE. tITP utilized optimum technique, achieving 19- 127-fold increase signal intensity when 200 ammonium formate (adjusted 4.00) leading BGE terminating electrolyte. Off-line eluate treatment procedures evaluated ensure compatibility selected preconcentration, process. Achieved recoveries were range 76-100% 12-54% 21-106% 25-98% saliva evaporated reconstituted solution achieve The validated across different matrices, offering good linearity, accuracy, precision, making it proteomic studies (e.g., therapeutic drug monitoring, biomarker research) samples.

Language: Английский

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Use of 3D printing to integrate microchip electrophoresis with amperometric detection

Analytical and Bioanalytical Chemistry, Journal Year: 2024, Volume and Issue: 416(21), P. 4749 - 4758

Published: April 6, 2024

Language: Английский

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Bipolar electrochemically generated fluorescence detector for microchip electrophoresis with and without a potentiostat: Application to reducible analyte detection

Sensors and Actuators Reports, Journal Year: 2025, Volume and Issue: 9, P. 100283 - 100283

Published: Jan. 14, 2025

Microchip electrophoresis (ME) is a separation method that makes it possible to quickly analyze multiple species in small volume samples miniaturized format. Two commonly employed detection approaches for ME are electrochemistry and fluorescence. Electrochemical can be direct of analytes via redox chemistry, while fluorescence generally offers lower limits but often requires analyte derivatization. The present study uses bipolar couple the two methods into single device as detector encompasses advantages both techniques. This system employs closed electrode (BPE) converts cathodic amperometric current produced at (sensing) pole response generated reporting pole. model analytes, benzoquinone resazurin, were separated detected using with reductive sensing BPE. A potentiostat was used control electrode. then converted through oxidation 2,7-dichlorodihydrofluorescein channel. subsequently modified run without by voltage bias BPE applied across channel generate electroosmotic flow well corresponding potentiostat-free mode evaluated determination 3-nitro-l-tyrosine, reducible biomarker oxidative stress. Lastly, sacrificial electrochemically active species, Trolox®, incorporated compensate background detection.

Language: Английский

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Advancement of microchip electrophoresis coupled with capacitively coupled contactless conductivity detection—from design to application (2018–2024)

Analytical Sciences, Journal Year: 2025, Volume and Issue: unknown

Published: April 25, 2025

Over the past three decades, microchip electrophoresis coupled with capacitively contactless conductivity detection (ME-C4D) has garnered considerable interest due to its various merits, including minimal sample consumption, compact structure, immediate detection, and high analytical precision. Continuous technological innovation improvement have significantly advanced ME-C4D in structural design, fabrication processes, experimental methodologies. As a result, application of this technology expanded into wider range electrochemical analysis fields, disease diagnosis, food safety assessment, environmental pollutant soil nutrient analysis. This review meticulously examines forefront over last five years. It methodically categorizes scrutinizes advancements from dimensions, newly emerged ME microchips, C4D electrodes, protocols, pioneering applications. Moreover, paper critically summarizes these developments, identifying prevailing limitations challenges within ME-C4D. Ultimately, it projects potential future trajectories for field ME-C4D, suggesting pathways overcome existing hurdles hinting at untapped possibilities that lie ahead.

Language: Английский

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Greenness of proteomic sample preparation and analysis techniques for biopharmaceuticals

TrAC Trends in Analytical Chemistry, Journal Year: 2023, Volume and Issue: 171, P. 117490 - 117490

Published: Dec. 19, 2023

Proteins are efficient therapeutic agents and attractive potential new biomarkers due to their physiological pathological functions. Further, biopharmaceuticals feature complex protein architectures that must be assayed quality controlled. Capillary electrophoresis (CE) has become one of the most powerful tools for proteomic analysis. CE is an eco-friendlier alternative traditional liquid chromatography; however, its true greenness environmental impact not very often confirmed by green metrics. Here, we assessed bottom-up, middle-up, top-down sample preparation analytical approaches analysis proteins using AGREEprep AGREE metric systems, respectively. The combined use both can help identify weaknesses overall procedure point toward greener alternatives. results showed trends in workflows should focused mainly on miniaturization devices applying in-line on-line preparation.

Language: Английский

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Enhancing affinity purification of monoclonal antibodies from human serum for subsequent CZE-MS analysis

Journal of Chromatography B, Journal Year: 2024, Volume and Issue: 1234, P. 123974 - 123974

Published: Jan. 8, 2024

Language: Английский

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