Noncovalently bilayer-coated capillaries for efficient and reproducible analysis of proteins by capillary electrophoresis.

摘要: The suitability of noncovalently bilayer-coated capillaries for the analysis proteins by capillary electrophoresis (CE) at medium pH was investigated. Fused-silica were coated simply successively flushing with a polybrene (PB) and poly(vinyl sulfonate) (PVS) solution. A protein test mixture used to evaluate performance capillaries. Comparisons bare fused-silica made. Several background electrolytes (BGEs) tested in combination PB-PVS coating, showing that optimum obtained using high BGE concentrations. With 300 mM Tris phosphate buffer (pH 7.0), good plate numbers (150,000-300,000), symmetrical peaks, favorable migration-time repeatabilities (RSDs below 0.8%) proteins. Using capillaries, peaks significantly broadened RSDs often exceeded 5%. It is concluded coating effectively minimizes adverse adsorption provides very stable electroosmotic flow (EOF). We also investigated potential commercially available bilayer (CEofix) analysis. demonstrated this peak symmetries can be achieved when CEofix ("accelerator") replaced common such as sodium or phosphate. Apparently, negatively charged polymer present "accelerator" interacts causing band broadening. utility coatings further illustrated separation interferon-alpha 2b, myoglobin carbonic anhydrase, degraded insulin sample time, profiling glycoprotein ovalbumin. In addition, it even presence concentrations human serum albumin up 60 mg/mL, still reproducible separations performance.