[Isolation of a HBV-PreS2 epitope from a random peptide library displayed on the bacterial flagellin].
OBJECTIVE
To establish an improved procedure for isolation and identification of epitopes from a random peptide library displayed on the bacterial surface.


METHODS
Epitopes 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.


RESULTS
Clones 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.


CONCLUSIONS
It's easy and quick to drive epitopes from a random peptide library displayed on the bacterial surface.
Xin
Zhong-tao
ZT
Department of Biochemistry, Institute of Basic Medical Sciences, Academy of Military Medical Science, Beijing 100850, China.
Xue
Yan-ning
YN
Gao
Ya-ping
YP
Mao
Can-quan
CQ
Liu
Chuan
C
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D001426:Bacterial Proteins / Q000276:immunology; D005408:Flagellin / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D016056:Immunodominant Epitopes / Q000276:immunology; D008969:Molecular Sequence Data; D019151:Peptide Library; D011498:Protein Precursors / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000956:Antigens, Viral; D001426:Bacterial Proteins; D006514:Hepatitis B Surface Antigens; D016056:Immunodominant Epitopes; D019151:Peptide Library; D011498:Protein Precursors; C050807:presurface protein 2, hepatitis B surface antigen; D005408:Flagellin