TrAC Trends in Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 118212 - 118212
Published: Feb. 1, 2025
Language: Английский
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
TrAC Trends in Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 118212 - 118212
Published: Feb. 1, 2025
Language: Английский
Citations
0