A novel approach for purification of recombinant proteins using the dextran-binding domain.
Using the dextran-binding domain (DBD) of a type of glucosyltransferase (GTF) from Streptococcus sobrinus, we have developed a novel method for purifying recombinant proteins. DBD-tagged green and red fluorescent proteins as well as the parent GTF and DBD moiety were adsorbed well to commercially available cross-linked dextran (such as Sephadex beads and Sephacryl beads), and eluted efficiently with water-soluble dextran. The purity of the eluted proteins after this one-step affinity purification was approximately 90% or better. The results suggest that DBD can be used as a powerful carrier for purification of various recombinant proteins.
Kaseda
K
K
Gene Discovery Research Center, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan. kaseda@nair.go.jp
Kodama
T
T
Fukui
K
K
Hirose
K
K
D000180:Acrylic Resins / Q000737:chemistry / Q000378:metabolism; D002846:Chromatography, Affinity / Q000191:economics / Q000379:methods; D003911:Dextrans / Q000737:chemistry / Q000378:metabolism*; D016695:Glycosyltransferases / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D008970:Molecular Weight; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D016986:Streptococcus sobrinus; D000097573:Red Fluorescent Protein
D000180:Acrylic Resins; D003911:Dextrans; D016695:Glycosyltransferases; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein; C053132:sephacryl S 500; C025614:sephadex