title: Continuous desalting of refolded protein solution improves capturing in ion exchange chromatography: A seamless process.
abstract: Truly continuous biomanufacturing processes enable an uninterrupted feed stream throughout the whole production without the need for holding tanks. We have utilized microporous anion and cation exchangers into which only salts, but not proteins, can penetrate into the pores for desalting of protein solutions, while diafiltration or dilution is usually employed for feed adjustments. Anion exchange and cation exchange chromatography columns were connected in series to remove both anions and cations. To increase operation performance, a continuous process was developed comprised of four columns. Continuous mode was achieved by staggered cycle operation, where one set of columns, consisting of one anion exchange and one cation exchange column, was loaded during the regeneration of the second set. Refolding, desalting and subsequent ion exchange capturing with a scFv as the model protein was demonstrated. The refolding solution was successfully desalted resulting in a consistent conductivity below 0.5 mS/cm from initial values of 10 to 11 mS/cm. With continuous operation process time could be reduced by 39% while productivity was increased to 163% compared to batch operation. Desalting of the protein solution resulted in up to 7-fold higher binding capacities in the subsequent ion exchange capture step with conventional protein binding resins.
authors.affiliation: Austrian Centre of Industrial Biotechnology, Vienna, Austria.
authors.forename: Nicole
authors.initials: N
authors.lastname: Walch
authors.affiliation: Austrian Centre of Industrial Biotechnology, Vienna, Austria.|Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
authors.forename: Alois
authors.initials: A
authors.lastname: Jungbauer
mesh_terms: D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D058849:Protein Refolding; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D002411:Cation Exchange Resins; D057127:Single-Chain Antibodies
keywords: Integrated biomanufacturing; Macroporous; Microporous; Single chain variable fragment; Staggered cycle operation