HPLC Analysis of mAb Charge Variants on TSKgel® CM-STAT
Materials
analytical column
CONDITIONS
column
TSKgel CM-STAT, 10 cm x 4.6 mm I.D., 7 μm particles (821966)
mobile phase
[A] 20 mM MES buffer, pH 6.0; [B] 0.5 M NaCl in buffer A, pH 6.0
gradient
0% B (0 min), 30% B (15 min), 100% B (15 min),
0% B (17 min), 10% B (17 min), 10% B (21 min)
flow rate
1 mL/min
column temp.
ambient
detector
UV, 280 nm
injection
20 μL
sample
monoclonal antibodies (mAb A through E)
Description
Analysis Note
A TSKgel CM-STAT weak cation exchange (WCX) column was applied to separate charge variants of several monoclonal antibodies. The typical analysis time on conventional 25 cm long WCX columns of about eighty minutes could be significantly reduced when separation was performed on a 10 cm TSKgel CM-STAT column, filled with 7 μm particles. The analysis profiles for five antibodies show that high resolution analysis can be obtained in about 20 minutes analysis time.
Citation
From "Ion Exchange Chromatography for the Characterization of Biotherapeutics" in The Supelco Reporter, Vol. 29.3, page 20.
Legal Information
TSKgel is a registered trademark of Tosoh Corporation