We explore the impact of two different stationary phases and ion-pair reagents on the retention behavior of a therapeutic peptide using reversed-phase liquid chromatography. This information is of fundamental importance for the development of reliable, selective, and fast analytical methods able to separate and identify the target peptide.
Single-column (batch) chromatography, involving two or more successive single-column (batch) chromatographic steps, is a standard approach for purifying biopharmaceuticals. Step one, known as the capture step, is used to remove product-related impurities, and step two, the polishing step, is used to remove product-related impurities. Here we present and illustrate the advantages of continuous chromatography for these separations: capture simulated moving bed (captureSMB) for the capture step and multicolumn countercurrent solvent gradient purification (MCSGP) for polishing.