This application note describes the design, optimization, and performance of BioResolve CX pH concentrates and their corresponding mobile phase system.
A new pH gradient mobile phase system, based on BioResolve CX pH Concentrates, has been developed to provide universally applicable cation-exchange separations of mAbs having a wide range of isoelectric point (pI) values. The composition and concentrations of buffer salts, as well as the pH range, were optimized using theory and empirical observations to achieve robust and high-resolution separation of mAbs. The performance of this method has been confirmed to be reproducible from batch-to-batch and to afford column lifetimes up to and beyond 500 injections. Moreover, it has been found that it is particularly advantageous to employ these mobile phase concentrates with BioResolve SCX mAb Columns. Method implementation and optimization has been simplified while both throughput and resolution have been improved. The BioResolve CX pH Concentrates, together with BioResolve SCX mAb Columns, provide an effective and efficient approach to mAb charge variant analysis.