[Construction and screening of hepatitis E virus-specific phage antibody combinatorial library].
OBJECTIVE
To construct HEV-specific phage combinatorial anti-body library and screen anti-HEV antibodies with neutralizing activity from the library.


METHODS
The total RNA was extracted from B-lymphocytes of 6 HE patients. Kappa chain and Fd segment of IgG gene were amplified respectively by RT-PCR using a set of Fab-specific primers. The amplified gene were inserted successively into vector pComb3 and electrotransformed E. coli XLI-Blue cells. Furthermore, the recombinant phage was rescued by being concultured with helper phage VCSM13 to construct HEV-specific phage anti-body library.


RESULTS
Fab displayed on the surface a as fusion protein with the N terminal of coat protein III, and 1. 8 x 10(7) clone library was established. Specific antibodies to HEV ORF2 recombinant antigen were acquired after five rounds of panning with HEV ORF2 recombinant antigen including neutralizing epitope.


CONCLUSIONS
Four clones exhibited specific binding to HEV ORF2 recombinant antigen including neutralizing epitope is identified by ELISA. The results show that we have got the recombinant phage antibodies.
Wei
Hua
H
Genetic Research Center, Medical School, Southeast University, Nanjing 210009, China. wei.xie@seu.edu.cn
Zhang
Jian-Qiong
JQ
Lu
Hai-Qin
HQ
Meng
Ji-Hong
JH
Lu
Xiao-Xuan
XX
Xie
Wei
W
D006508:Hepatitis Antibodies / Q000235:genetics*; D016752:Hepatitis E virus / Q000276:immunology*; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D014764:Viral Proteins / Q000276:immunology*
D006508:Hepatitis Antibodies; D007145:Immunoglobulin kappa-Chains; C400971:ORF2 protein, Hepatitis E virus; D019151:Peptide Library; D014764:Viral Proteins