Continuous separation of protein loaded nanoparticles by simulated moving bed chromatography.
Bottom Line: In the case of beta casein where no multimers are present we achieved 89% purity and 90% recovery of loaded nanoparticles in the Raffinate and an extract free of particles (92% purity).Using a tangential flow filtration unit with 5kDa cutoff membrane we proved that the extract can be concentrated for recycling of protein and buffer.The calculated space-time-yield for loaded nanoparticles was 0.25g of loaded nanoparticles per hour and liter of used resin.
Affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna (BOKU), Muthgasse 18, 1190 Vienna, Austria.Show MeSH
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Mentions: To prove the applicability of TFF for concentration of protein and reuse of buffer we collected the extract of the SMB–SEC separation of 70 nm silica nanoparticles and beta-casein and concentrated it over the course of 80 min (Fig. 7). We achieved a total concentration factor of 2.8, which allows this solution to be used again for loading of nanoparticles, reaching a yield of 96%. We also proved that the resulting buffer is free of protein and ready to be used in the SMB system again (Fig. 7).
Affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna (BOKU), Muthgasse 18, 1190 Vienna, Austria.