Isolation of a HBV-PreS2 epitope from a random peptide library displayed on the bacterial flagellin.
- Author:
Zhong-tao XIN
1
;
Yan-ning XUE
;
Ya-ping GAO
;
Can-quan MAO
;
Chuan LIU
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Animals; Antigens, Viral; immunology; Bacterial Proteins; immunology; Flagellin; immunology; Hepatitis B Surface Antigens; genetics; immunology; isolation & purification; Immunodominant Epitopes; immunology; Molecular Sequence Data; Peptide Library; Protein Precursors; genetics; immunology; isolation & purification
- From: Acta Academiae Medicinae Sinicae 2003;25(1):56-59
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish an improved procedure for isolation and identification of epitopes from a random peptide library displayed on the bacterial surface.
METHODSEpitopes were screened from FliTrx random peptide library by a monoclonal antibody 3B9 against HBV-PreS2 protein. The enrichment was monitored in each round. Higher affinity clones were obtained by increasing the washing strength and randomly selected for sequencing and Western blot analysis.
RESULTSClones specifically binding to antibody were enriched in each round. Ten sequences were obtained from sixteen sequenced clones, seven of them contained the common motif RXRGXY with high homogeneity to 135-140 amio acids in HBV-PreS protein and have positive results in Western blot analysis. The other three sequences have no typical motif RXRGXY and showed different Western blot results.
CONCLUSIONSIt's easy and quick to drive epitopes from a random peptide library displayed on the bacterial surface.
