Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary...

Area of Science:

• Analytical Chemistry
• Separation Science
• Materials Science

Background:

• Capillary electrophoresis (CE) often faces challenges with basic protein adsorption onto silica capillary walls.
• Existing coatings may not provide sufficient stability or reproducibility for complex protein separations.

Purpose of the Study:

• To synthesize and evaluate cationized hydroxyethylcellulose (cat-HEC) as a novel physically adsorbed capillary coating for CE.
• To assess the performance of cat-HEC in separating basic proteins compared to unmodified hydroxyethylcellulose (HEC).

Main Methods:

• Synthesis of cationized hydroxyethylcellulose (cat-HEC) in the laboratory.
• Application of cat-HEC as a physically adsorbed coating in CE capillaries via simple flushing.
• Comparative analysis of protein separation with and without cat-HEC coating.
• Evaluation of coating stability and reproducibility in consecutive runs.

Main Results:

• The cat-HEC coating effectively suppressed the adsorption of basic proteins onto the silica capillary wall.
• Efficient basic protein separations with excellent reproducibility were achieved using the cat-HEC coating.
• The positively charged cat-HEC coating induced an anodal electroosmotic flow (EOF).
• Long-term stability of the cat-HEC coating was demonstrated for high-throughput separations.

Conclusions:

• Cationized hydroxyethylcellulose (cat-HEC) is a simple, effective, and stable capillary coating for CE.
• Cat-HEC enables reproducible separation of basic proteins by minimizing undesirable adsorption.
• This novel coating is suitable for high-throughput protein analysis in capillary electrophoresis.