| Autor: |
Boumaiza M; Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Université Tunis El Manar, Tunis, Tunisia., Trabelsi K; Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Université Tunis El Manar, Tunis, Tunisia.; Life Science Department, Biotechnology Programme, College of Graduate Studies, Arabian Gulf University, Manama, Bahrain., Choucha Z; Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Université Tunis El Manar, Tunis, Tunisia., Akrouti I; Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Université Tunis El Manar, Tunis, Tunisia., Leone S; Department of Chemical Sciences, University of Naples Federico II, Naples, Italy., Picone D; Department of Chemical Sciences, University of Naples Federico II, Naples, Italy., Kallel H; Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Université Tunis El Manar, Tunis, Tunisia.; UnivercellsVaccines, Nivelles, Belgium. |
| Abstrakt: |
Hepatitis E virus (HEV) is a nonenveloped virus causing an emerging zoonotic disease posing a severe threat to the public health in the world, especially to pregnant women. In this study, a truncated form (aa 368-606) of the open reading frame 2 of the capsid protein ( t ORF2-HEV), a major structural protein of HEV, was expressed in Escherichia coli . This work characterizes for the first time, the fused Glutathione-S-Transferase-tagged t ORF2 (GST- t ORF2) and t ORF2-HEV forms in E. coli . The fusion protein was purified by affinity chromatography with a purity higher than 90% and to yield about 27% after thrombin digestion. The purified GST- t ORF2 protein was then characterized by western blot, using anti-GST antibodies, and CD spectroscopy. The GST- t ORF2 and t ORF2-HEV proteins were shown to be efficient to develop an ELISA test to detect anti-HEV IgG in mice sera immunized with a recombinant full length ORF2 protein. Sera showed a significant increase of the absorbance signal at 450 nm, in plate wells coated with a quantity of 0.5, 1 and 2 µg of proteins. ELISA plates coated with the purified GST- t ORF2 and t ORF2-HEV showed similar response when compared to the HEV ELISA where total insect cell lysate, infected with the recombinant baculovirus expressing full ORF2, was used as positive control. |
