Immunogenic efficacy of differently produced recombinant vaccines candidates against Pseudomonas aeruginosa infections.
Three different variants of the recombinant hybrid outer membrane protein OprF (aa 190-342)-OprI (aa 21-83) could be obtained in high yield after expression in Escherichia coli. The hybrid protein was modified N terminally, either with a minimal histidine tag or with a homologous sequence of OprF. Both recombinant proteins were purified by nickel chelate affinity chromatography under native and denaturing conditions, and this produced three suitable candidates for a vaccination trial, protein His-F-I, which was purified in its native as well as in its refolded form; and the native purified N terminally extended protein, ex-F-I. In mice, significantly higher antibody titers and survival rates after challenge with Pseudomonas aeruginosa were observed following immunization with protein His-F-I, purified under native conditions.
von Specht
B U
BU
Chirurgische Universitätsklinik der Universität Freiburg, Chirurgische Forschung, Hugstetter Strasse 55, 79106, Freiburg, Germany. specht@ch11.ukl.uni-freiburg.de
Gabelsberger
J
J
Knapp
B
B
Hundt
E
E
Schmidt-Pilger
H
H
Bauernsachs
S
S
Lenz
U
U
Domdey
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005982:Glutathione Transferase / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D018272:Porins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011552:Pseudomonas Infections / Q000276:immunology / Q000517:prevention & control*; D011550:Pseudomonas aeruginosa / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000276:immunology*
D000907:Antibodies, Bacterial; D018272:Porins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D005982:Glutathione Transferase