Practical considerations for preparing polymerized phospholipid bilayer capillary coatings for protein separations.

Area of Science:

• Analytical Chemistry
• Materials Science

Background:

• Phosphorylcholine (PC) phospholipid bilayers are effective capillary coatings resisting protein adsorption.
• Limited long-term chemical and physical stability hinder widespread use of PC coatings.

Purpose of the Study:

• Investigate polymerized lipid bilayer capillary coatings for improved stability and separation performance.
• Optimize coating conditions by examining pH and capillary inner diameter effects.

Main Methods:

• Utilized polymerized synthetic phospholipids for fused silica capillary coatings.
• Evaluated separation performance, reproducibility (run-to-run, day-to-day, column-to-column), and long-term stability.
• Tested coating performance across a range of pH values and capillary inner diameters (≤50 μm and wider).

Main Results:

• Polymerized phospholipid coatings demonstrated enhanced stability and reproducibility.
• Coatings maintained stability for protein separations across a broad pH range (4.0-9.3).
• Smaller inner diameter capillaries (≤50 μm) showed significantly improved coating stability and reproducibility.

Conclusions:

• Polymerized phospholipid coatings offer a stable and reproducible solution for capillary-based separations.
• The wide pH stability range and improved performance in smaller capillaries present significant advantages for analytical applications.
• Optimized capillary size is crucial for maximizing the benefits of polymerized phospholipid coatings.