Investigation and optimization of elution buffer effects on the elution profile for the cation exchange chromatography resin POROS XS : In association with Cobra Biologics

Abstract: The goal of this project was to investigate how elution buffer properties affectthe protein elution in the polishing purification step with a cation exchange chromatographyconsisting of POROS XS resin. Hence be able to better predict thebehavior of the resin, two model proteins were used for this investigation (K100and Y102). This project included lab experiments (small scale), and design ofexperiments using MODDE software to evaluate the optimal conditions. Thefactors pH, conductivity, sample load and buffer concentration were investigatedon antibody Y102 while only pH and conductivity were investigated as factorsfor antibody K100. The evaluated responses were the elution volume, aggregateamount and the amount of monomeric antibody (target protein). The results indicatedthat the elution behaviour for POROS XS resin was antibody-dependentand not general for the resin. The responses obtained different models for thetwo antibodies investigated. However, since the aggregate and monomeric proteinfor K100 had curvature indications more experiments should be performedto establish the different behaviour of aggregate and monomer for the respectiveproteins.