Journal
JOURNAL OF BIOTECHNOLOGY
Volume 83, Issue 1-2, Pages 3-12Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0168-1656(00)00311-4
Keywords
Pseudomonas aeruginosa; outer membrane protein; vaccine
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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. (C) 2000 Elsevier Science B.V. All rights reserved.
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