A method for increasing the stability and uniformity of a PEGylated G-CSF
polypeptide having at least one PEG moiety attached to the epsilon amino
group of a lysine residue or the N-terminal amino group and at least one
PEG moiety attached to a hydroxyl group, comprising subjecting the
polypeptide to an elevated pH of above 8.0 for a period of time suitable
to remove PEG moieties attached to a hydroxyl group, and reducing the pH
to about 8.0 or lower; as well as PEGylated G-CSF polypeptides and
compositions produced according to the method and methods for increasing
neutrophil levels in a patient using the PEGylated G-CSF polypeptides and
compositions.