HPLC Analysis of Size Exclusion Separation of Proteins and PEGylated Proteins

HPLC Analysis of Size Exclusion Separation of Proteins and PEGylated Proteins

Conditions

column Zenix SEC-150, 30 cm x 7.8 mm I.D., 3 µm particles, 150 Ã…, (Z777018)
column temp. Ambient
mobile phase 150 mM phosphate buffer, pH 7.0
flow rate 1.0 mL/min
sample 1. Protein with MW 70KD (from an undisclosed drug development company) 2. Protein 1 attached with 1 PEG (MW 40KD) molecule 3, Protein 1 attached with 2 PEG (MW 40KD) molecules
injection 20 µL, 2.0 mg/mL
detector UV, 214 nm

Description

Analysis Note Pegylation of biotherapeutic drugs improves solubility, increases half life in the blood stream and reduces immunogenicity. Size exclusion chromatography (SEC) is often applied to characterize protein–polyethylene glycol (PEG) conjugates and establish the number of attached PEG chains, since SEC is a precise and robust technique that combines ease of set up and strainghtforward method develpopment.
Featured Industry Life Science and Biopharma
suitability application for HPLC

Materials

     
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