title: Dialysis strategies for protein refolding: preparative streptavidin production.
abstract: We have investigated different dialysis strategies for the refolding of recombinant streptavidin, and present a novel dialysis setup featuring gradual dilution dialysis and continuous protein feeding into a dialysis sack. A denaturing dialysis buffer is exchanged gradually by dilution with refolding buffer and it is demonstrated that the refolding yield can be increased from 45 to 75% by lowering the dilution rate. In addition, continuous feeding of protein to the dialysis sack increases the yield by 5 to 10%. The principle of gradual dilution dialysis is amenable to stringent regulation and we suggest it to be applied for other insoluble protein targets.
authors.affiliation: Laboratory of BioDesign, Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 10 C, DK-8000, Aarhus C, Denmark.
authors.forename: Hans Peter
authors.initials: HP
authors.lastname: Sørensen
authors.forename: Hans Uffe
authors.initials: HU
authors.lastname: Sperling-Petersen
authors.forename: Kim Kusk
authors.initials: KK
authors.lastname: Mortensen
mesh_terms: D000465:Algorithms; D002850:Chromatography, Gel; D003956:Dialysis / Q000295:instrumentation / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D019791:Guanidine / Q000737:chemistry; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D011489:Protein Denaturation; D017510:Protein Folding; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D019809:Streptavidin / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D019809:Streptavidin; D019791:Guanidine