[Construction and preliminary application of a human natural phage antibody library with diversity].
OBJECTIVE
To construct a human natural phage single-chain antibody (scFv) library with diversity.


METHODS
V(H) and V(L) genes were amplified by RT-PCR and hemi-PCR from peripheral blood lymphocytes of healthy persons. The V genes were assembled to form scFv by overlap PCR and cloned into phagemid pCANTAB-5E, and then transformed into E. coli TG1 by electroporation to construct a human natural phage scFv library. The diversity and gene family of antibody gene were analysed by sequencing and the specific antibodies against various antigens were screened through bio-panning.


RESULTS
A human natural phage scFv library with diversity and 2x10(8) sink size was constructed successfully. The specific human scFvs against 5 antigens were obtained by bio-panning.


CONCLUSIONS
A human natural phage scFv library with diversity is constructed successfully and can be applied to human antibody preparation.
Wang
Jin
J
The Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, Xiamen University, Research Center for Medical Molecular Virology of Fujian Province, Xiamen 361005, China. wxluo@jingxian.xmu.edu.cn
Luo
Wen-Xin
WX
Li
Li-Feng
LF
Chen
Ying-Wei
YW
Wang
Ming-Qiao
MQ
Zhang
Jun
J
Xia
Ning-Shao
NS
D000906:Antibodies / Q000032:analysis / Q000235:genetics* / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000032:analysis / Q000235:genetics / Q000276:immunology; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D000906:Antibodies; D000941:Antigens; D007135:Immunoglobulin Variable Region; D019151:Peptide Library