[Screening and identification of mimic epitopes of monoclonal antibodies against hantaan virus using phage display technique].
OBJECTIVE
To obtain mimic epitopes of monoclonal antibody (mAb) against Hantaan virus, and identify preliminarily their immunological characteristics.


METHODS
Group-specific mAbs F3 and B11 were used as selective molecules for biopanning. After biopanning, the positive phages were identified by ELISA and DNA sequencing. Among the positive phages, F3-8, B11-18, B11-20 and B11-24 were selected to immunize BALB/c mice three times, respectively. The antiserum was then titered and confirmed by sandwich ELISA and competition ELISA.


RESULTS
After 3 to 4 rounds of effective screening, the majority of the selected clones were found able to react to mAb F3 or B11 in a dose-dependent manner, but not to BSA, rHBsAg or other unrelated mAbs. The amino acid sequences of the clones binding to mAb F3 contained an identical sequence MHGPTKNQMWHT, which had higher homology to 750-759 amino acids between a pair of cysteines within glycoprotein G2 of HTNV/SEOV, while those of clones binding to mAb B11 had no evident homologous regions within HTNV/SEOV proteins. The specific antibodies increased significantly following immunization with phage peptides, which indicated that the phage-displayed peptides had not only good antigenicity, but also strong immunoreactivity.


CONCLUSIONS
The phage-displayed peptides could mimic the epitope of HFRSV antigen, which would provide the potential for preparing more effective epitope-based vaccines and specific diagnostic reagents.
Wang
Chang-jun
CJ
Institute of Military Medicine, Nanjing Command, Nanjing 210002, China. science2008@yahoo.com.cn
Tang
Jia-qi
JQ
Tao
Kai-hua
KH
Li
Xian-fu
XF
Bai
Wei
W
Guo
Heng-bing
HB
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000914:Antibodies, Viral / Q000276:immunology* / Q000378:metabolism; D018604:Epitope Mapping / Q000379:methods; D000939:Epitopes; D018063:Hantaan virus / Q000235:genetics / Q000276:immunology*; D007106:Immune Sera; D007114:Immunization; D018716:Molecular Mimicry; D019151:Peptide Library; D017386:Sequence Homology, Amino Acid
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D007106:Immune Sera; D019151:Peptide Library