EGG-WHITE PROTEIN SEPARATION DEVELOPMENT - EFFECT OF BUFFERS AND UNTREATED FUSED-SILICA VS NEUTRAL COVALENTLY BONDED COLUMNS

A method for the separation of three egg white proteins, lysozyme. conalbumin, and ovalbumin, was developed based on desired criteria of reproducible baseline separation of all components in under 25 minutes. Fixed parameters were 50cm effective column length, 200 volts/cm field strength. Sodium phosphate, sodium borate. and phosphate/borate combination buffers were compared in different ionic strengths. The performance of bonded phase columns, CElect(Tm)-P1 and CElect-H (patent pending) were compared to that of untreated fused silica. The optimal separation was obtained using 100mM phosphate / 60 mM borate buffer, pH 8.0 on the hydrophilic CElect-P1 bonded phase column.