Continuous matrix-assisted refolding of proteins

摘要: A refolding reactor was developed for continuous matrix-assisted of proteins. The composed an annular chromatography system and ultrafiltration to recycle aggregated proteins produced during the reaction. feed solution containing denatured protein continuously fed rotating bed perfused with buffer promoting folding protein. As passed through column, it separated from chaotropic reducing agents process took place. Native aggregates could be due different molecular size. exit stream collected, concentrated by recycled solution. high concentrations in enabled dissociation consequently were again reactor. When initial mixture is used up, only buffer-containing are fully converted native This resulted a stoichiometric conversion its correctly folded state. tested bovine α-lactalbumin as model Superdex 75 PrepGrade size-exclusion medium. yield 30% active monomer batch improved 41% at recycling rate 65%. Assuming that can redissolved into quantitative manner, close 100% possible. method also applied other chromatographic principles suited separation aggregates.