[Preparation of human single chain Fv antibody specifically against hepatitis C virus E2 antigen].
OBJECTIVE
To identify human single chain Fv antibody (ScFv) against hepatitis C viral E2 antigen and its value clinically.


METHODS
The recombinant phages were panned by E2 antigen which was coated in a microtiter plate. After five rounds of biopanning, 56 phage clones were identified specific to E2 antigen. The affinity and specificity of ScFv were evaluated by ELISA and immunohistochemistry, respectively.


RESULTS
The data of E2-ScFv DNA digestion and DNA sequencing showed that the ScFv gene was composed of 750bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against HCV E2 antigen had a specific combination character with hepatitis C virus E2 antigen.


CONCLUSIONS
ScFv, having a sutestantial affinity and specificity and being easy to prepare, is valuable in the detection of HCV E2 antigen.
Zhong
Yanwei
Y
Gene Therapy Research Centre, Institute of Infectious Diseases, 302nd Hospital of PLA, Beijing 100039, China.
Cheng
Jun
J
Shi
Shuangshuang
S
Wang
Gang
G
Xia
Xiaobing
X
Tian
Xiaojun
X
Li
Li
L
Zhang
Lingxia
L
Chen
Jumei
J
D000595:Amino Acid Sequence; D000915:Antibody Affinity; D000918:Antibody Specificity; D001483:Base Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D014759:Viral Envelope Proteins / Q000276:immunology*
D007128:Immunoglobulin Fragments; D014759:Viral Envelope Proteins; C091015:glycoprotein E2, Hepatitis C virus