Highly hydrophilic group-substituted celluloses were immobilized on the fused silica capillary inner wall for capillary isoelectric focusing (CIEF) of proteins. The resultant capillaries were evaluated under typical CIEF conditions, and the results were compared with those obtained by using polyvinyl alcohol (PVA)- and linear polyacrylamide (LPA)-coated capillaries. A minimal resolved pI difference of 0.013 was obtained with a carrier ampholyte (pH 3-10) for hydroxyethyl cellulose- and hydroxypropyl cellulose-coated capillaries, while 0.019 was obtained on PVA- and LPA coated capillaries. Changing substituted groups from hydroxyl to hydroxylmethyl decreased the CIEF resolution from ~2.5 to ~1.0 for tested proteins at mobilization speed of ~0.9 cm min-1. Little loss in CIEF resolution was observed when increasing capillary length from 30 to 150 cm, however, flat peaks were observed when increasing column diameter up to >100 um. The present work indicates that mobilization speed of
Revised: May 30, 2002 |
Published: March 1, 2000
Citation
Shen Y., and R.D. Smith. 2000.High-Resolution Capillary Isoelectric Focusing of Proteins Using Highly Hydrophilic-Substituted Cellulose-Coated Capillaries.Journal of Microcolumn Separations 12, no. 3:135-141.PNNL-SA-32141.