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: 70 nm silica nanoparticles loaded with BSA or beta-casein were separated from free protein (Fig. 4) by Sephacryl300 batch experiment. Protein loaded nanoparticles passed through the column and were eluted in the void fraction, but the protein was retained. The differences in the peak height of eluted nanoparticles loaded with different proteins are explained by the different molar extinction coefficients of the protein as well as the different amount of adsorbed protein. We did not achieve base line separation for BSA and protein loaded nanoparticles, but SMB does not require base line separation to yield pure fractions of a binary mixture .
Affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna (BOKU), Muthgasse 18, 1190 Vienna, Austria.