Influence of polyelectrolyte capillary coating conditions on protein analysis in CE.
CE of biomolecules is limited by analyte adsorption on the capillary wall. To prevent this, monolayer or successive multiple ionic-polymer layers (SMILs) of highly charged polyelectrolytes can be physically adsorbed on the inner capillary surface. Although these coatings have become commonly used in CE, no systematic investigation of their performance under different coating conditions has been carried out so far. In a previous study (Nehmé, R., Perrin, C., Cottet, H., Blanchin, M. D., Fabre, H., Electrophoresis 2008, 29, 3013-3023), we investigated the influence of different experimental parameters on coating stability, repeatability and peptide peak efficiency. Optimal coating conditions for monolayer and multilayer (SMILs) poly(diallyldimethylammonium) chloride/ poly(sodium 4-styrenesulfonate) coated capillaries were determined. In this study, the influence of polyelectrolyte concentration and ionic strength of the coating solutions, and the number of coating layers on coating stability and performance in limiting protein adsorption was carried out. EOF magnitude and repeatability were used to monitor coating stability. Coating ability to limit protein adsorption was investigated by monitoring variations of migration times, time-corrected peak areas and separation efficiency of test proteins. The separation performance of polyelectrolyte coatings were compared with those obtained with bare silica capillaries.
Nehmé
Reine
R
Institut des Biomolécules Max Mousseron, UMR, Université Montpellier 1, Université Montpellier 2, CNRS, Montpellier, France.
Perrin
Catherine
C
Cottet
Hervé
H
Blanchin
Marie-Dominique
MD
Fabre
Huguette
H
D000327:Adsorption; D004573:Electrolytes / Q000737:chemistry*; D053841:Electroosmosis; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D009994:Osmolar Concentration; D011095:Polyethylenes / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D015203:Reproducibility of Results; D013451:Sulfonic Acids / Q000737:chemistry
D004573:Electrolytes; D011095:Polyethylenes; D011108:Polymers; D011506:Proteins; D000644:Quaternary Ammonium Compounds; D013451:Sulfonic Acids; C041004:poly-N,N-dimethyl-N,N-diallylammonium chloride; C077114:styrenesulfonic acid polymer