title: Antigenicity of a recombinant NS3 protein representative of ATPase/helicase domain from hepatitis C virus.
abstract: It has been shown that the Hepatitis C virus nonstructural NS3 protein possesses at least two enzymatic domains: a serine-protease domain and an adenosine triphosphatase (ATPase)/helicase domain. In this report, a truncated fragment of NS3 (26 kDa), representing main epitopes from the (ATPase)/helicase domain, has been expressed in Escherichia coli. The recombinant protein was purified by Ion Metal Affinity Chromatography (IMAC) with more than 90% purity. The recognition of B-cell linear epitopes in the NS3 protein was evaluated by immunoblot. The recombinant NS3 protein was reduced and carboxymethylated, and the recognition of either conformational and/or linear B-cell determinants was evaluated by ELISA. The inclusion of the recombinant NS3 protein in a third-generation diagnostic system UltraMicroELISA (UMELISA) allowed an increase in the sensitivity, due to the detection of a new variety of false-negative sera in blood donor test samples.
authors.affiliation: Diagnostics Production Plant, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba.
authors.forename: N
authors.initials: N
authors.lastname: Pentón
authors.forename: A
authors.initials: A
authors.lastname: Musacchio
authors.forename: J M
authors.initials: JM
authors.lastname: Rivera
authors.forename: J
authors.initials: J
authors.lastname: Roca
authors.forename: M
authors.initials: M
authors.lastname: Ponce
authors.forename: D
authors.initials: D
authors.lastname: Rodríguez
authors.forename: A
authors.initials: A
authors.lastname: Caballero
authors.forename: Y I
authors.initials: YI
authors.lastname: Tallo
authors.forename: R E
authors.initials: RE
authors.lastname: Narciandi
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D004926:Escherichia coli / Q000378:metabolism; D005285:Fermentation; D015151:Immunoblotting; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000251:Adenosine Triphosphatases; D004265:DNA Helicases; D000939:Epitopes; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases