Production and characterization of a fusion form of hepatitis E virus tORF2 capsid protein in Escherichia coli. Issue 6 (1st July 2021)

• Record Type: Journal Article
• Authors: Boumaiza, Mohamed; Trabelsi, Khaled; Choucha, Zeineb; Akrouti, Ines; Leone, Serena; Picone, Delia; Kallel, Héla
• Is Part Of: Preparative biochemistry & biotechnology. Volume 51:Issue 6(2021)
• Page Start: 562
• Page End: 569
• Publication Date: 2021-07-01
• Subjects: CD spectroscopy -- ELISA -- GST-tORF2 fusion protein -- Hepatitis E virus -- tORF2-HEV; Biochemistry -- Technique -- Periodicals; Biotechnology -- Technique -- Periodicals; Biochemistry -- methods -- Periodicals; Biotechnology -- methods -- Periodicals; 660.6
• DOI: 10.1080/10826068.2020.1836656
• Languages: English
• ISSNs: 1082-6068

Abstract

Abstract: 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.