21034775
not annotated - annotated - LINNAEUS only
Cross-genotypic polyclonal anti-HCV antibodies from human ascitic fluid.
Many anti-HCV antibodies are available, but more are needed for research and clinical applications. This study examines whether ascitic fluid from cirrhotic patients could be a source of reagent-grade antibodies. Ascitic fluid from 29 HCV patients was screened by ELISA for anti-HCV antibodies against three viral proteins: core, NS4B, and NS5A. Significant patient-to-patient variability in anti-HCV antibody titers was observed. Total ascitic fluid IgG purified by Protein-A chromatography reacted with HCV proteins in immunoblots, cell extracts, and replicon-expressing cells. Affinity-purification using synthetic peptides as bait allowed the preparation of cross-genotypic antibodies directed against pre-selected regions of HCV core, NS4B, and NS5A proteins. The performance of the polyclonal antibodies was comparable to that of monoclonal antibodies. Anti-NS4B antibody preparations reacted with genotype 1a, 1b, and 2a NS4B proteins in immunoblots and allowed NS4B to be localized in replicon-expressing cells. Ascitic fluid is an abundant source of human polyclonal cross-genotypic antibodies that can be used as an alternative to blood. This study shows the utility of selectively purifying human polyclonal antibodies from ascitic fluid. Affinity purification allows antibodies to be selected that are comparable to monoclonal antibodies in their ability to react with targeted regions of viral proteins.
Ann file
T1 Out-of-scope 32 35 HCV
N1 Reference T1 Taxonomy:11103
T2 Species 52 57 human
N2 Reference T2 Taxonomy:9606
T3 Out-of-scope 84 87 HCV
N3 Reference T3 Taxonomy:11103
T4 Species 309 312 HCV
N4 Reference T4 Taxonomy:11103
T5 Out-of-scope 353 356 HCV
N5 Reference T5 Taxonomy:11103
T6 Out-of-scope 471 474 HCV
N6 Reference T6 Taxonomy:11103
T7 Species 579 582 HCV
N7 Reference T7 Taxonomy:11103
T8 Species 1133 1138 human
N8 Reference T8 Taxonomy:9606
T9 Species 1276 1281 human
N9 Reference T9 Taxonomy:9606
T10 Species 804 807 HCV
N10 Reference T10 Taxonomy:11103 Hepacivirus C
#1 AnnotatorNotes T10 TODO check