URG7 is an anti-apoptotic protein which consists of 99 amino acid residues up regulated by antigen x during the HBV infection. The first 74 amino acids are identical to those of the multidrug resistance protein 6 (MRP6), while the amino acid residues from 75 to 99 are specific for URG7 protein. Immuno-informatics tools and secondary structure analysis were carried out to identify the antigenic properties of this URG7 sequence. The 75-99 peptide was synthesized by the solid-phase method, structurally characterized by CD spectroscopy and conjugated to a protein carrier. New Zealand white rabbits were immunized and sera were tested for anti-peptide specific antibodies by ELISA and western blot analysis. Finally ELISA test with human sera was performed. Rabbits immunized with the 75-99 peptide produce antibodies that recognize both the 75-99 peptide and the URG7 recombinant polypeptide. Moreover, both antigens allowed for the detection of the anti-URG7 antibodies in sera of healthy and HBV infected subjects although with a different discriminant threshold. Our data suggested that peptide ELISA assay against the specific sequence of the URG7 protein allows with good sensitivity and specificity for the detection of anti-URG7 antibodies in sera from HBV infected patients.

Immunochemical Characterization of the Specific Sequence of URG7 Protein

DI NARDO, Elvira;
2016

Abstract

URG7 is an anti-apoptotic protein which consists of 99 amino acid residues up regulated by antigen x during the HBV infection. The first 74 amino acids are identical to those of the multidrug resistance protein 6 (MRP6), while the amino acid residues from 75 to 99 are specific for URG7 protein. Immuno-informatics tools and secondary structure analysis were carried out to identify the antigenic properties of this URG7 sequence. The 75-99 peptide was synthesized by the solid-phase method, structurally characterized by CD spectroscopy and conjugated to a protein carrier. New Zealand white rabbits were immunized and sera were tested for anti-peptide specific antibodies by ELISA and western blot analysis. Finally ELISA test with human sera was performed. Rabbits immunized with the 75-99 peptide produce antibodies that recognize both the 75-99 peptide and the URG7 recombinant polypeptide. Moreover, both antigens allowed for the detection of the anti-URG7 antibodies in sera of healthy and HBV infected subjects although with a different discriminant threshold. Our data suggested that peptide ELISA assay against the specific sequence of the URG7 protein allows with good sensitivity and specificity for the detection of anti-URG7 antibodies in sera from HBV infected patients.
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ELISA; Immunogenic peptide; Hepatitis B virus; Preneoplastic marker; URG7
Di Nardo E.; Ostuni A.; Cuviello F; Salvia AM; D'Auria F; Statuto T; Miglionico R; Carretta V; Musto P; Bisaccia F
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2318/1616080
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