Document Detail


Identification of B cell epitopes of hepatitis C virus RNA dependent RNA polymerase.
MedLine Citation:
PMID:  12020787     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The aim of this study was to identify the B cell epitopes of hepatitis C virus (HCV) NS5B RNA dependent RNA polymerase (RdRp). The truncated HCV NS5B protein NS5B-dc21 was expressed in Escherichia coli and its antigenicity was confirmed by Enzyme-Linked Immunosorbent Assay (ELISA) using 130 HCV-positive human sera and 15 negative sera. Antibodies specific to NS5B-dc21 protein were purified by affinity chromatography using sepharose-4B coupled with the recombinant protein. A 12-mer phage displayed random peptide library was screened four rounds with the purified antibodies. Three epitopes were identified from the phage library, which correspond to amino acids 2444-2452, 2521-2528, and 2915-2925 of HCV RdRp. These epitopes were then expressed in E. coli as fusion proteins with phage M13 pIII protein. ELISA demonstrated that two of these epitopes (P4 and P34, corresponding to amino acids 2443-2452 and amino acids 2512-2528, respectively) have good reactivity and sensitivity. Mutagenesis study of P4 peptide showed that this epitope, which is derived from a phage displayed library, exhibited higher affinity with HCV serum than the corresponding original HCV sequences.
Authors:
Lihua Hou; Guixin Du; Yigang Tong; Haitao Wang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of virological methods     Volume:  104     ISSN:  0166-0934     ISO Abbreviation:  J. Virol. Methods     Publication Date:  2002 Jun 
Date Detail:
Created Date:  2002-05-21     Completed Date:  2002-08-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8005839     Medline TA:  J Virol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1-8     Citation Subset:  IM    
Affiliation:
Department of Applied Molecular Biology, Institute of Microbiology and Epidemiology, No. 20 Dongdajie Street, Fengtai District, 100071, Beijing, PR China. houlihua@hotmail.com
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Antibodies, Viral / immunology,  isolation & purification
Epitope Mapping
Epitopes, B-Lymphocyte / genetics,  immunology*
Hepacivirus / enzymology*
Molecular Sequence Data
Peptide Library
RNA Replicase / genetics,  immunology*
Viral Nonstructural Proteins / genetics,  immunology*
Chemical
Reg. No./Substance:
0/Antibodies, Viral; 0/Epitopes, B-Lymphocyte; 0/NS-5 protein, hepatitis C virus; 0/Peptide Library; 0/Viral Nonstructural Proteins; EC 2.7.7.48/RNA Replicase

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