Recent stationary phase-based fractionation strategies in proteomic analysis

Area of Science:

• Proteomics
• Analytical Chemistry
• Chromatography

Background:

• Proteomic analysis relies on effective sample prefractionation to enhance depth and coverage.
• Stationary phase-assisted techniques are crucial for separating complex biological samples.
• Recent advancements focus on improving prefractionation efficiency prior to mass spectrometry.

Purpose of the Study:

• To review stationary phase-assisted sample prefractionation techniques in proteomic analysis.
• To categorize applications based on retention modes used before liquid chromatography-tandem mass spectrometry.
• To highlight novel online prefractionation platforms and instrumental configurations.

Main Methods:

• Literature review of proteomic analysis articles published between 2019 and 2022.
• Grouping prefractionation applications by chromatographic retention mode.
• Analysis of online and offline instrumental configurations for sample preparation.
• Focus on techniques preceding low-pH reversed-phase liquid chromatography-tandem mass spectrometry.

Main Results:

• Affinity chromatography was the most frequently used mode for sample fractionation.
• The popularity order of chromatographic modes was: Affinity > Size Exclusion > Hydrophilic Interaction > High-pH Reversed-Phase > Ion Exchange.
• Novel online prefractionation platforms show significant promise for streamlined workflows.
• Low-pH reversed-phase liquid chromatography-tandem mass spectrometry remains a common final analysis step.

Conclusions:

• Stationary phase-assisted prefractionation is vital for advancing proteomic analysis.
• Affinity chromatography leads in popularity for sample fractionation in recent studies.
• The development of online platforms offers opportunities for more efficient proteomic workflows.
• Continued innovation in prefractionation techniques is essential for deeper proteome coverage.