Incorporation of hydrophobic selectivity in capillary electrophoresis: analysis of recombinant insulin-like growth factor I variants.
A highly selective electrophoretic system employing differential hydrophobic interaction was evaluated for the quantitative determination of recombinant insulin-like growth factor I (IGF-I) variants. The system consisted of mixed aqueous-organic buffers containing suitable amounts of a zwitterionic detergent. In addition, a neutral hydrophilic coating was attached to the wall of the capillary to minimize analyte adsorption and electroosmotic flow. The zwitterionic detergent acted as a hydrophobic selector, allowing independent optimization of the electrophoretic and hydrophobic selectivities in the separation system. The extent of hydrophobic interaction was conveniently adjusted by varying the type and amount of organic modifier. Complete resolution of a mixture of IGF-I variants with closely related mass-to-charge ratios was achieved. Quantitative analysis of IGF-I process samples agreed well with HPLC results. Finally, the approach was found to be compatible with on-line capillary electrophoresis-mass spectrometry.
Nashabeh
W
W
Barnett Institute, Northeastern University, Boston, Massachusetts 02115.
Greve
K F
KF
Kirby
D
D
Foret
F
F
Karger
B L
BL
Reifsnyder
D H
DH
Builder
S E
SE
D000595:Amino Acid Sequence; D004586:Electrophoresis; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000032:analysis*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000032:analysis
D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I