Monoclonal antibodies (mAbs) have been widely applied in the therapeutic area, which requires large amounts of the product and high purity, which is achieved only by chromatographic methods. Among them, hydrophobic interaction chromatography (HIC) exploits the hydrophobicity characteristics of mAbs through the interaction between non-polar regions present on their surface and hydrophobic ligands. However, one of the major problems in the development of these processes is the time taken to obtain high quantities of product. As an alternative, the continuous mode can be used, in which the multi-column process is executed, as in Periodic Chromatography Counter-Current (PCCC), resulting in high productivity with increased utilization of useful column capacity. Thus, this project aims to evaluate the effect of residence time, salt concentration and salt type on the adsorption of Trastuzumab in hydrophobic interaction adsorbents (Toyopearl Phenyl-650 and Phenyl Sepharose 6 Fast Flow) for its purification in a process 3C-PCCC. Trastuzumab adsorption experiments will be carried out in batch mode with subsequent characterization of heterogeneity using low angle X-ray scattering (SAXS) and differential scanning calorimetry (DSC) techniques. Then, rupture curves will be obtained to determine the dynamic binding capacity (DBC) as a function of residence time. For the purposes of selectivity and purity, SDS-PAGE and ELISA will be performed. Knowing the time of the batch experiment and the cycle time for each continuous system, the productivity gain will be evaluated considering how much antibody is purified per hour in each of the operation modes.
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