Optimizing protein separations with cation exchange chromatography using Agilent Buffer Advisor

Optimizing protein separations with cation exchange chromatography using Agilent Buffer Advisor Protein separation with the Agilent 1260 Infinity Bio‐inert Quaternary LC System

This Technical Overview shows that the Agilent Buffer Advisor software in com‐ bination with the Agilent 1260 Infinity Bio‐inert Quaternary LC System is an ideal solution for automated protein separation by ionic strength gradients. Usually, pH scouting using premixed two‐component gradients is time‐consuming and work‐ intensive. Dynamically mixed four‐component gradients calculated by the Buff...

Optimizing Protein Separations with Agilent Weak Cation-Exchange Columns

Columns containing weak cation-exchange stationary phases offer a degree of versatility when used for separation of positively charged proteins. Of particular interest, is the difference in selectivity that can be obtained by adjusting the pH of the mobile phase eluent. We explore the parameters of interest and how these can be changed to optimize the resolution of the compounds of interest usi...

pH Gradient Elution for Improved Separation of Monoclonal Antibody Charge Variants

Ion exchange is a very useful technique to separate protein mixtures under mild conditions, most commonly by applying a shallow gradient of increasing salt concentration as eluent. pH gradient elution is used much less frequently, except in specialist applications such as chromatofocusing, and can require complex buffer systems. However, today’s modern quaternary HPLC systems are ideally suited...

Faster Separations Using Agilent Weak Cation Exchange Columns

Systematic generation of buffer systems for pH gradient ion exchange chromatography and their application.

pH gradient protein separations are widely used techniques in the field of protein analytics, of which isoelectric focusing is the most well known application. The chromatographic variant, based on the formation of pH gradients in ion exchange columns is only rarely applied due to the difficulties to form controllable, linear pH gradients over a broad pH range. This work describes a method for ...