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Monoclonal antibodies dowstream processing using continous hydrophobic interaction chromatography

Grant number: 18/16533-1
Support Opportunities:Scholarships abroad - Research
Start date: February 01, 2019
End date: January 31, 2020
Field of knowledge:Engineering - Chemical Engineering - Industrial Operations and Equipment for Chemical Engineering
Principal Investigator:Iara Rocha Antunes Pereira Bresolin
Grantee:Iara Rocha Antunes Pereira Bresolin
Host Investigator: Alois Jungbauer
Host Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil
Institution abroad: Universität für Bodenkultur Wien, Austria  

Abstract

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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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ANTUNES PEREIRA BRESOLIN, IARA ROCHA; LINGG, NICO; LAZZAROTTO BRESOLIN, IGOR TADEU; JUNGBAUER, ALOIS. Hydrophobic interaction chromatography as polishing step enables obtaining ultra-pure recombinant antibodies. Journal of Biotechnology, v. 324, n. S, . (18/16533-1)