Sensitive method to analyse free PEG-maleimide

摘要

The present invention relates generally to a novel method using HPLC and fluorescence detection of free PEG-mal in PEGylated proteins and PEG-mal raw materials by adding a fluorescent label to the free PEG-mal.

主权项

1. A method to determine the concentration of polyethylene glycol-maleimide (PEG-mal) in a PEG-mal solution said method comprising the steps of
(a) adding the fluorescent labeled peptide probe consisting of SEQ ID NO: 1, to said solution; (b) allowing the fluorescent labeled peptide probe in the solution of step (a) to react under dark conditions with the PEG-mal present in the solution to provide fluorescent labeled PEG-mal; (c) separating by fractionation the fluorescent labeled PEG-mal from the free polyethylene glycol using high performance liquid chromatography (HPLC) wherein the fluorescent fraction identifies the PEG-mal.