RESUMO
Hepatitis delta virus (HDV), a recently discovered infectious agent, participates in severe, often lethal forms of acute and chronic hepatitis and liver cirrhosis. Based on theoretical analysis of secondary structure, hydrophilicity and acrophilicity data, several regions of HDV antigen, presumably containing B-epitopes, have been revealed and the corresponding peptides have been synthesized by the solid phase method. All the peptides obtained reacted with the respective antipeptide rabbit sera. The peptides and their conjugates with BSA or KLH were used for ELISA with individual and pooled anti-HD-positive sera from patients with chronic delta hepatitis. The high antigenicity of the peptide 65-80 shows that one of the antigenically active regions of HDAg is situated between these amino acid residues and that the peptide may be used for detection of anti-HD antibodies in patients blood sera.
Assuntos
Capsídeo/metabolismo , Vírus Delta da Hepatite/metabolismo , Peptídeos/síntese química , Proteínas do Core Viral/metabolismo , Sequência de Aminoácidos , Animais , Antígenos Virais/imunologia , Ensaio de Imunoadsorção Enzimática , Vírus Delta da Hepatite/imunologia , Humanos , Imunização , Técnicas Imunoenzimáticas , Masculino , Dados de Sequência Molecular , Peptídeos/imunologia , CoelhosRESUMO
We synthesized the 24-41, 30-36, 31-36, 24-30 fragments of the preS1-region of the hepatitis B (subtype ayw) envelope. The peptides were prepared by the solid phase synthesis on perfluorpolyethylene polymer grafted with polystyrene. The peptide chains were elongated from C-terminus using activated esters and symmetrical anhydrides of Boc-amino acids, cleaved off the solid phase by HBr or TFMSA in TFA, purified by gel filtration, and, after conjugation with protein carriers, inoculated into test animals. The resultant antibodies were shown to react with peptides. The blood sera from patients with acute hepatitis B reacted with the conjugates of peptides 24-41, 30-36, 31-36 in the immunoenzymic solid phase assay. The monoclonal antibodies for the preS1-polypeptide were shown to react with peptides 24-41, 30-36, 31-36 and with their conjugates. The results obtained were proved by the data of the epitope-mapping with overlapping hexapeptides.
Assuntos
Epitopos/metabolismo , Hepatite B/metabolismo , Peptídeos/síntese química , Proteínas do Envelope Viral/metabolismo , Sequência de Aminoácidos , Cromatografia em Gel , Hepatite B/imunologia , Técnicas Imunoenzimáticas , Dados de Sequência Molecular , Proteínas do Envelope Viral/imunologiaAssuntos
Antígenos Virais/imunologia , Capsídeo/imunologia , Vírus Delta da Hepatite/imunologia , Fragmentos de Peptídeos/imunologia , Proteínas do Core Viral/imunologia , Antígenos Virais/síntese química , Hepatite D/imunologia , Humanos , Epitopos Imunodominantes/imunologia , Fragmentos de Peptídeos/síntese química , Conformação Proteica , Relação Estrutura-AtividadeRESUMO
We have synthesized the 24-41 fragment of the preS region of the hepatitis B (subtype ayw) envelope. The peptide was prepared by the solid phase synthesis on perfluoropoly-ethylene polymer grafted with polystyrene. The peptide chain was elongated from C-terminus using pentafluorophenyl- and p-nitrophenyl esters of Boc-amino acids. The peptide was cleaved off the solid phase by HBr in CF3COOH, purified by gel filtration, and, after conjugation with serum albumin (BSA), inoculated into mice. The resultant antibodies were shown to react with the peptide. The blood sera from patients with acute hepatitis B reacted with the peptide-BSA conjugates in the immunoenzymic solid phase assay.