HPLC Analysis of PEG on Two Zenix® SEC-300 Columns in Tandem
Materials
analytical column
Zenix® SEC-300 Gel Filtration Column, 3 μm
L × I.D. 30 cm × 7.8 mm, 3 μm particle size, stainless steelguard cartridge
Zenix® SEC-300 Gel Filtration Guard Column, 3 μm
L × I.D. 5 cm × 7.8 mm, 3 μm particle size, stainless steelCONDITIONS
column
Zenix SEC-300 (2 ea), 30 cm x 7.8 mm I.D., 3 μm partcles, 300 Å, connected in tandem (Z777033)
mobile phase
150 mM Sodium Phosphate Buffer pH=7.0
flow rate
0.5 mL/min
column temp.
25 °C
detector
Refractive Index at 30 °C
injection
10 μL
sample
1.PEG 81,400 Da (2mg/mL)
2.PEG 50,630 Da (2mg/mL)
3.PEG 40,000 Da (2mg/mL)
4.PEG 26,100 Da (2mg/mL)
5.PEG 8,730 Da (2mg/mL)
Description
General description
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.
This chromatogram shows the separation of four polyethyleneglycol polymers varying in molecular mass 8730 to 81,400 Da on two Zenix® SEC-300 columns in series.
Legal Information
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