The hepatitis A virus (HAV) capsid protein VP1, VP2 and VP3 are exposed at the virion surface and should therefore contain antigenic determinants. Algorithms for hydrophilicity, antigenicity and flexibility were used to predict probable antigenic sites. Synthesis of 7- to 23-membered overlapping peptides from seven sites, viz.
View Article and Find Full Text PDFA modified enzyme immunoassay based on adsorption of antihepatitis A virus (HAV) IgG-HRPO conjugate and monoclonal antibodies to HAV were used to investigate antigenic differences between mature HAV virions and subviral particles with different buoyant densities in CsCl produced in HAV-infected cells. The mature virions (1.34 g/cm3) appeared to have common antigenic determinants with subviral particles (1.
View Article and Find Full Text PDFPeptide (P6) representing amino acids 62-75 of HAV capsid protein VP3 was synthesized. Pure P6, octamer form of P6, and P6 conjugated to KLH were injected into rabbits. The sera against these preparations reacted with denaturized VP3 in immunoblotting tests; however, only anti-P6-KLH serum was capable of binding the native HAV.
View Article and Find Full Text PDFThe Western blot procedure with highly specific antipeptide antibody was applied to identify the electrophoretic mobility of hepatitis A virus capsid proteins. Polypeptides with molecular weights of 33, 29 and 27 kDa proved to be VP1, VP2, and VP3 proteins as they reacted with sera generated to VP1 recombinant protein and to synthetic oligopeptides 42-62 VP2 and 62-75 VP3, respectively.
View Article and Find Full Text PDFHepatitis A virus (HAV) was propagated in continuous rhesus monkey embryo kidney cells (FRhK-4 line). HAV RNA extracted with phenol from purified HAV pretreated with proteinase K was used for molecular cloning experiments. Two radioactive plasmids, pHAV-23 and pHAV-5p, containing HAV cDNA fragments complementary to structural and nonstructural parts of HAV genome, respectively, were found to be useful for discrimination between mature virions and subviral structures.
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