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Recombinant HpaA purified from Escherichia coli has biological properties similar to those of native Helicobacter pylori HpaA.
- Source :
-
APMIS : acta pathologica, microbiologica, et immunologica Scandinavica [APMIS] 2003 Mar; Vol. 111 (3), pp. 389-97. - Publication Year :
- 2003
-
Abstract
- The aim of this study was to recombinantly produce and purify Helicobacter pylori adhesin A (HpaA) from Escherichia coli and compare it to purified native H. pylori HpaA, for potential use as a vaccine antigen. The hpaA gene was cloned from H. pylori, transferred to two different expression vectors, and transformed into E. coli. Expression of rHpaA was analysed by immunoblot, inhibition ELISA, and semi-quantitative dot-blot. Using affinity chromatography, rHpaA was purified from E. coli and native HpaA from H. pylori. The binding of both purified proteins to sialic acid was analysed and antibody titres to native and rHpaA were compared after intraperitoneal immunisation of C57/Bl mice. The rHpaA protein was highly expressed in E. coli from both vectors. Purified recombinant and native HpaA bound similarly to fetuin but also to the non-sialylated asialofetuin. Both native HpaA and rHpaA induced comparable amounts of specific antibodies in serum after immunisation and they were identical in double immunodiffusion. In conclusion, rHpaA was successfully produced in E. coli. Purified rHpaA showed biological properties similar to those of native HpaA isolated from H. pylori and may therefore be further used as an antigen in the development of a vaccine against H. pylori infection.
- Subjects :
- Adhesins, Bacterial genetics
Adhesins, Bacterial pharmacology
Animals
Antibodies, Bacterial biosynthesis
Antibodies, Bacterial blood
Antibodies, Monoclonal immunology
Blotting, Western
Chromatography, Affinity
Cloning, Molecular
DNA, Bacterial chemistry
DNA, Bacterial genetics
Enzyme-Linked Immunosorbent Assay
Escherichia coli chemistry
Escherichia coli genetics
Escherichia coli metabolism
Female
Helicobacter pylori genetics
Helicobacter pylori immunology
Immunization
Immunodiffusion
Mice
Mice, Inbred C57BL
N-Acetylneuraminic Acid metabolism
Recombinant Proteins genetics
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Recombinant Proteins pharmacology
alpha-Fetoproteins immunology
alpha-Fetoproteins metabolism
Adhesins, Bacterial immunology
Adhesins, Bacterial metabolism
Helicobacter pylori metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0903-4641
- Volume :
- 111
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- APMIS : acta pathologica, microbiologica, et immunologica Scandinavica
- Publication Type :
- Academic Journal
- Accession number :
- 12752218
- Full Text :
- https://doi.org/10.1034/j.1600-0463.2003.t01-1-1110203.x