Model-based description of peptide retention on doped reversed-phase media

Autor: Gianluca Buffolino, Nicola Forrer, Massimo Morbidelli, Rushd Khalaf, Alessandro Butté
Rok vydání: 2015
Předmět:
Zdroj: Journal of chromatography. A. 1407
ISSN: 1873-3778
Popis: Reversed-phase (RP) chromatography is one of the main tools for the preparative purification of therapeutic peptides. In previous works [1] , [2] , a new type of RP chromatography, doped reversed-phase chromatography (DRP) was presented. By adding small amounts (up to 15% of the surface ligands) of repulsive ion exchange ligands to a traditional RP material, significant improvements in peptide purification performance were observed, at the same or in similar operating conditions. These improvements included increases in selectivity in diluted conditions (up to twice as high), increases in yield in preparative conditions (up to 20% higher) and in productivity in preparative conditions (up to twice as high), when compared to RP materials [2] . A proper physical model is developed in this work to quantitatively explain and rationalize this behavior. The developed model is then used to correctly fit the retention data of several peptides in different buffering conditions. The increase in selectivity is related to a controlled decrease in free surface area available for adsorption due to the ionic ligands creating a repulsive sphere the analytes cannot enter. This decrease in adsorption surface is calculated using Debye–Huckel theory, and in combination with linear solvent strength theory, allows for the quantitative description of peptide retention on DRP media.
Databáze: OpenAIRE