[Preparation and characteristics of the human  single-chain Fv antibodies to NS5B].
OBJECTIVE
To prepare and identify the human  single-chain Fv (scFv) antibody to hepatitis C virus(HCV) NS5B.


METHODS
After   the peripheral blood mononuclear cells were collected from HCV patients,  the single-chain antibody Fv library was constructed by amplifying  the whole variable region of heavy chain(V(H);) and variable region of light chain(V(L);).   Targeting  NS5B,  the scFv gene was  cloned into pET16b, and  then transfected  into BL21 cells. The scFv was induced to  express in the cells.  Sequencing and analysis of the scFv were performed.


RESULTS
The single-chain antibody Fv library was successfully constructed; two scFvs with higher binding activity were expressed;  the sequencing result showed that   the scFv  was  consistent with the expectation.


CONCLUSIONS
Preparation of the human  scFv  antibody to HCV NS5B provides  a basis for immunotherapy to HCV.
Department of Haematology, Tangdu Hospital, Fourth Military Medical University, Xi'an, China. chen-renan@163.com
Ren-an
RA
Chen
Jiang-hong
JH
Tian
Hua
H
Wang
D000595:Amino Acid Sequence; D000915:Antibody Affinity; D001483:Base Sequence; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000276:immunology; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase
D057127:Single-Chain Antibodies; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase