Toward Unrivaled Chromatographic Resolving Power in Proteomics: Design and Development of Comprehensive Spatial Three-Dimensional Liquid-Phase Separation Technology

被引:2
作者
Eeltink, Sebastiaan [1 ]
De Vos, Jelle [1 ,2 ]
Desmet, Gert [1 ]
机构
[1] Vrije Univ Brussel, Dept Chem Engn, Brussels, Belgium
[2] RIC Grp, Kortrijk, Belgium
关键词
multidimensional separations; three-dimensional liquid chromatography; 3D-LC; peak capacity; top-down proteomics; protein analysis; biomarker discovery; PEAK-CAPACITY; MICROFLUIDIC DEVICE; RESOLUTION; LASER; OPTIMIZATION; PEPTIDES; ISOFORMS;
D O I
10.1146/annurev-anchem-061522-044510
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Spatial comprehensive three-dimensional chromatography (3D-LC) offers an innovative approach to achieve unprecedented resolving power in terms of peak capacity and sample throughput. This advanced technique separates components within a 3D separation space, where orthogonal retention mechanisms are incorporated. The parallel development of the second- and third-dimension stages effectively overcomes the inherent limitation of conventional multidimensional approaches, where sampled fractions are analyzed sequentially. This review focuses on the design aspects of the microchip for spatial 3D-LC and the selection of orthogonal separation modes to enable the analysis of intact proteins. The design considerations for the flow distributor and channel layout are discussed, along with various approaches to confine the flow during the subsequent development stages. Additionally, the integration of stationary phases into the microchip is addressed, and interfacing to mass spectrometry detection is discussed. According to Pareto optimality, the integration of isoelectric focusing, size-exclusion chromatography, and reversed-phase chromatography in a spatial 3D-LC approach is predicted to achieve an exceptional peak capacity of over 30,000 within a 1-h analysis, setting a new benchmark in chromatographic performance.
引用
收藏
页码:475 / 493
页数:19
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