title: Constructing Chimeric Antigen for Precise Screening of HTLV-I Infection.
abstract: Individual preparation of two human T-cell lymphotropic virus type I (HTLV-I) diagnostic GST fused peptides (MTA-1 and GD21) is time-consuming and expensive. The aim of this study was to design a novel single chimeric antigen (SCA) to obviate separate expression of proteins and reduce the cost of reagent preparation. Structural protein fragments, including immunodominant B cell linear epitopes, were selected and different SCAs were designed. Tertiary structure, epitope exposure, solubility and stability were calculated for each SCA and compared with each other. The synthetic DNA encoding the interested SCA was sub-cloned into pET32a expression vector, expressed as a soluble form in Escherichia coli BL21 (DE3) cells and purified under native condition using affinity chromatography. The SDS-PAGE results indicated that thioredoxin-fused SCA was successfully expressed as a soluble form in E. coli BL21 (DE3) cells. The results of ELISA confirmed that SCA reacted with anti-HTLV-I antibodies in a concentration-dependent manner. Our results indicated that the designed SCA may be a good candidate for the screening of HTLV-I carriers with antigen-antibody-based tests.
authors.affiliation: Department of Clinical Biochemistry, Faculty of Medicine, Sabzevar University of Medical Science, Sabzevar, Iran.
authors.forename: Hafez
authors.initials: H
authors.lastname: Heydari Zarnagh
authors.affiliation: Department of Pediatrics, Faculty of Medicine, Sabzevar University of Medical Science, Sabzevar, Iran.
authors.forename: Kazem
authors.initials: K
authors.lastname: Hassanpour
authors.affiliation: Department of Biotechnology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Rasaee
mesh_terms: D000956:Antigens, Viral / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D015490:HTLV-I Infections / Q000175:diagnosis*; D015368:Human T-lymphotropic virus 1 / Q000276:immunology*; D006801:Humans; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012995:Solubility
chemical_list: D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins
keywords: Antigenicity; Chimeric protein; ELISA; HTLV-I