Abstract
The analysis of a recombinant granulocyte macrophage colony stimulating factor (GM-CSF) dosage form by free solution capillary electrophoresis (FSCE), capillary isoelectric focusing (cIEF), and high-performance liquid chromatography (HPLC) is described. The quantitative use of capillary electrophoresis, whether FSCE or cIEF, will always be prone to special problems, such as sensitivity to differences in salt concentrations between the standard and sample, and can not match the ruggedness of HPLC. The usable quantitative linear range for both HPLC and FSCE surpass that achieved for cIEF methods by a factor of 10 or greater. The FSCE system, utilizing an octyl bonded/Brij-35 coated capillary, did not work for all proteins examined. This is probably due to an interaction of the protein with the bonded phase or the absorbed Brij-35. In contrast, the cIEF method worked well for all proteins tested thus far, yielding high efficiency and resolution comparable to slab gel isoelectric focusing. This paper addresses the potential for using free solution capillary electrophoresis and capillary isoelectric focusing as a quantitative analytical tool. Also, the effect of salt in the dosage form on quantitation, reproducibility, and efficiency of capillary electrophoresis methods is also discussed.
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