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Objective: To describe the expression and immunogenic efficacy of differently produced recombinant vaccines candidates against Pseudomonas aeruginosa infections. Methods: The hybrid protein OprF (aa 190-342)-Opr I (aa 21-83) 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. Results :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 P. aeruginosa were observed, following immunization with protein His-F- I purified under native conditions. Conclusion: A hybrid OprF-Opr I molecule was cloned and a purification method which yields a protective vaccine against P. aeruginosa infections was establis

来源:第二军医大学学报 1998 年 19卷 z1期

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来源:
第二军医大学学报 1998 年 19卷 z1期
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Pseudomonas aeruginosa outer membrane protein vaccine
Objective: To describe the expression and immunogenic efficacy of differently produced recombinant vaccines candidates against Pseudomonas aeruginosa infections. Methods: The hybrid protein OprF (aa 190-342)-Opr I (aa 21-83) 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. Results :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 P. aeruginosa were observed, following immunization with protein His-F- I purified under native conditions. Conclusion: A hybrid OprF-Opr I molecule was cloned and a purification method which yields a protective vaccine against P. aeruginosa infections was establis