Capillary electrophoretic separation of poly(ethylene glycol)-modified granulocyte-colony stimulating factor.
We evaluated the utility of capillary electrophoretic methods for analyzing poly(ethylene glycol) (PEG)-modified granulocyte-colony stimulating factor (G-CSF), a long-acting form of GCSF for the treatment of cancer therapy-induced neutropenia. Low- and high-molecularweight PEG-G-CSF conjugates prepared with aldehyde-activated PEG-5K and PEG-20K were separated by high-performance size-exclusion chromatography (HP-SEC), capillary zone electrophoresis (CZE), and sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE). HPSEC showed low resolution for separating mono- and di-PEG-G-CSFs. SDS-CGE had higher resolution, but required a long analysis and had low peak efficiency. CZE could successfully separate both PEG-5K- and PEG-20K-conjugated G-CSFs with a running time of 20 min and high peak efficiency. In conclusion, CZE was better than SDS-CGE for separating PEG-G-CSF conjugates and will be useful for PEGylation studies, such as reaction monitoring for optimization of the PEGylation reaction, and purity and stability tests of PEG-G-CSF.
Lee
Kyung Soo
KS
College of Pharmacy, Kyungsung University, Busan, 608-736, Korea.
Na
Dong Hee
DH
D002626:Chemistry, Pharmaceutical; D002850:Chromatography, Gel; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry / Q000302:isolation & purification*; D006801:Humans; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry / Q000302:isolation & purification*; D013678:Technology, Pharmaceutical / Q000379:methods*; D013997:Time Factors
C486981:pegylated granulocyte colony-stimulating factor, human; D016179:Granulocyte Colony-Stimulating Factor; D011092:Polyethylene Glycols