High-resolution capillary isoelectric focusing of proteins using highly hydrophilic-substituted cellulose-coated capillaries

✍ Scribed by Yufeng Shen; Richard D. Smith

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 121 KB
• Volume: 12
• Category: Article
• ISSN: 1040-7685
• DOI: 10.1002/(sici)1520-667x(2000)12:3<135::aid-mcs2>3.0.co;2-5

✦ Synopsis

Highly hydrophilic group-substituted celluloses were immobilized on Ž . 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 Ž . Ž . PVA -and linear polyacrylamide LPA -coated capillaries. A minimal resolved p I Ž . 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 ca. 2.5 to ca. 1.0 for tested proteins at mobilization speed of ca. 0.9 cm min y1 . 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 m. The present work indicates that mobilization speeds of -1 cm min y1 should be used to achieve high CIEF resolution.