HPLC Analysis of mAb Aggregates on TSKgel® UltraSW Aggregate: Effect of Mobile Phase Additives on Resolution
CONDITIONS
column
TSKgel UltraSW Aggregate 30 cm x 7.8 mm I.D., 3 μm particles (822856)
mobile phase
0.1 mol/L sodium phosphate buffer, pH 6.7, with an addition of either 0.2 mol/L arginine (A), or 0.2 mol/L proline (B)
flow rate
1.0 mL/min
column temp.
ambient
detector
UV, 280 nm
injection
20 μL
sample
mAB, aggregated at 75 °C for 5 min
Description
Analysis Note
While particle size, packing quality and column length are all important considerations in optimizing mAb aggregate analysis, mobile phase additives can also contribute to improving your separation. The addition of amino acids such as lysine and arginine to the mobile phase, increase aggregate recovery, while glutamine, proline and arginine improve monomer aggregate resolution, as well as monomer fragment resolution. Although the increases in resolution are not drastic, they confirm that increased resolution due to the use of an advanced mobile phase is possible and that mobile phase testing can contribute to a more reliable and robust aggregate analysis.
Legal Information
TSKgel is a registered trademark of Tosoh Corporation