title: Efficient purification of recombinant proteins fused to maltose-binding protein by mixed-mode chromatography.
abstract: Two mixed-mode resins were evaluated as an alternative to conventional affinity resins for the purification of recombinant proteins fused to maltose-binding protein (MPB). We purified recombinant MBP, MBP-LacZ and MBP-Leap2 from crude Escherichia coli extracts. Mixed-mode resins allowed the efficient purification of MBP-fused proteins. Indeed, the quantity of purified proteins was significantly higher with mixed-mode resins, and their purity was equivalent to that obtained with affinity resins. By using purified MBP, MBP-LacZ and MBP-Leap2, the dynamic binding capacity of mixed-mode resins was 5-fold higher than that of affinity resins. Moreover, the recovery for the three proteins studied was in the 50-60% range for affinity resins, and in the 80-85% range for mixed-mode resins. Mixed-mode resins thus represent a powerful alternative to the classical amylose or dextrin resins for the purification of recombinant proteins fused to maltose-binding protein.
authors.affiliation: EA 4135, ESTBB, Université V. Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux cedex, France.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Cabanne
authors.forename: Jérôme
authors.initials: J
authors.lastname: Pezzini
authors.forename: Gilles
authors.initials: G
authors.lastname: Joucla
authors.forename: Agnès
authors.initials: A
authors.lastname: Hocquellet
authors.forename: Caroline
authors.initials: C
authors.lastname: Barbot
authors.forename: Bertrand
authors.initials: B
authors.lastname: Garbay
authors.forename: Xavier
authors.initials: X
authors.lastname: Santarelli