Efficient purification of recombinant proteins fused to maltose-binding protein by mixed-mode chromatography.
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.
Cabanne
Charlotte
C
EA 4135, ESTBB, Université V. Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux cedex, France.
Pezzini
Jérôme
J
Joucla
Gilles
G
Hocquellet
Agnès
A
Barbot
Caroline
C
Garbay
Bertrand
B
Santarelli
Xavier
X