Purification of LamB proteins using continuous elution electrophoresis: a comparison with immunoaffinity chromatography.
LamB is a membrane protein that allows the exposition of a foreign peptide on the surface of a recombinant E. coli cells. An immunopurified hybrid LamB protein has been used to elicit high-titre antibodies to a foreign epitope. Looking for a simpler purification procedure we have compared the traditional approach, which includes affinity chromatography, to continuous elution electrophoresis, in the purification of two different hybrid LamB proteins as foreign epitopes. The results obtained showed that both methods yielded the same purification, although the electrophoretic procedure had a higher yield. Continuous-elution electrophoresis could be a useful tool for the purification of membrane proteins.
Flamminio
G
G
Institute of Microbiology, University of Brescia, Italy.
Monti
E
E
Gargiulo
F
F
varinacci
C
C
Magni
P
P
Martinelli
F
F
Caimi
L
L
Caruso
A
A
Preti
A
A
Turano
A
A
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002462:Cell Membrane / Q000648:ultrastructure; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli; D008961:Models, Structural; D008970:Molecular Weight; D010957:Plasmids; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins