title: Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies.
abstract: Computational tools are reliable alternatives to laborious work in chimeric protein design. In this study, a chimeric antigen was designed using computational techniques for simultaneous detection of anti-HTLV-I and anti-HBV in infected sera. Databases were searched for amino acid sequences of HBV/HLV-I diagnostic antigens. The immunodominant fragments were selected based on propensity scales. The diagnostic antigen was designed using these fragments. Secondary and tertiary structures were predicted and the B-cell epitopes were mapped on the surface of built model. The synthetic DNA coding antigen was sub-cloned into pGS21a expression vector. SDS-PAGE analysis showed that glutathione fused antigen was highly expressed in E. coli BL21 (DE3) cells. The recombinant antigen was purified by nickel affinity chromatography. ELISA results showed that soluble antigen could specifically react with the HTLV-I and HBV infected sera. This specific antigen could be used as suitable agent for antibody-antigen based screening tests and can help clinicians in order to perform quick and precise screening of the HBV and HTLV-I infections.
authors.affiliation: Department of Clinical Biochemistry, Faculity of Medical Science, Tarbiat Modares University, Tehran, Iran Hafez.heydari@modares.ac.ir.
authors.forename: Hafez
authors.initials: H
authors.lastname: Heydari Zarnagh
authors.affiliation: Department of Virology, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran ravanshad@modares.ac.ir.
authors.forename: Mehrdad
authors.initials: M
authors.lastname: Ravanshad
authors.affiliation: Department of Immunology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran pourfa@modares.ac.ir.
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Pourfatollah
authors.affiliation: Department of Biotechnology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran rasaee_m@modares.ac.ir.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Rasaee
mesh_terms: D000595:Amino Acid Sequence; D019295:Computational Biology / Q000379:methods*; D015480:Deltaretrovirus Antibodies / Q000032:analysis*; D015485:Deltaretrovirus Antigens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D006800:Deltaretrovirus Infections / Q000175:diagnosis; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000276:immunology; D006509:Hepatitis B / Q000175:diagnosis; D006510:Hepatitis B Antibodies / Q000032:analysis*; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis*; D014412:Tumor Virus Infections / Q000175:diagnosis
chemical_list: D015480:Deltaretrovirus Antibodies; D015485:Deltaretrovirus Antigens; D018985:Epitopes, B-Lymphocyte; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D016056:Immunodominant Epitopes; D011993:Recombinant Fusion Proteins