Open AccessImmunochemical Characterization of the Specific Sequence of URG7 Protein
Author(s) -
Angela Ostuni,
Flavia Cuviello,
Salvia Am,
Francesco D’Auria,
Teodora Statuto,
Di Nardo E,
Rocchina Miglionico,
Carretta,
P. Musto,
Faustino Bisaccia
Publication year - 2016
Publication title -
journal of biomolecular research and therapeutics
Language(s) - English
Resource type - Journals
ISSN - 2167-7956
DOI - 10.4172/2167-7956.1000146
Subject(s) - characterization (materials science) , sequence (biology) , computational biology , biology , biochemistry , materials science , nanotechnology
URG7 is an anti-apoptotic protein which consists of 99 amino acid residues up regulated by antigen x during the\udHBV infection. The first 74 amino acids are identical to those of the multidrug resistance protein 6 (MRP6), while the\udamino acid residues from 75 to 99 are specific for URG7 protein. Immuno-informatics tools and secondary structure\udanalysis were carried out to identify the antigenic properties of this URG7 sequence. The 75-99 peptide was\udsynthesized by the solid-phase method, structurally characterized by CD spectroscopy and conjugated to a protein\udcarrier. New Zealand white rabbits were immunized and sera were tested for anti-peptide specific antibodies by\udELISA and western blot analysis. Finally ELISA test with human sera was performed.\udRabbits immunized with the 75-99 peptide produce antibodies that recognize both the 75-99 peptide and the\udURG7 recombinant polypeptide. Moreover, both antigens allowed for the detection of the anti-URG7 antibodies in\udsera of healthy and HBV infected subjects although with a different discriminant threshold. Our data suggested that\udpeptide ELISA assay against the specific sequence of the URG7 protein allows with good sensitivity and specificity\udfor the detection of anti-URG7 antibodies in sera from HBV infected patients