期刊
JOURNAL OF CHROMATOGRAPHY A
卷 908, 期 1-2, 页码 235-241出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0021-9673(00)00739-1
关键词
hydrophobic interaction chromatography; preparative chromatography; proteins; poly(ethylene glycol); poly(ethylene glycol) bis-vinylsulfone
Cation- and anion-exchange chromatography can be used to purify a polyethylene glycol-linked protein dimer (PEG dimer) made with M-r 20 000 PEG bis-vinylsulfone, even when there are no net charge differences between the components that are being separated. The retention time on ion-exchange generally is inversely proportional to the PEG protein ratio (on a mass basis). One of the biggest challenges in developing the process for making this PEG dimer was the quality of the PEG linker. Reversed-phase HPLC can be used to determine both size heterogeneity and the degree of end-group activation of M-r 20 000 PEG bis-vinylsulfone. In addition, we have found that hydrophobic interaction chromatography can be used make more size homogeneous preparations of M-r 20000 PEG bis-vinylsulfone, which significantly increased the recovery of the PEG dimer. (C) 2001 Elsevier Science B.V. All rights reserved.
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