[Construction of a human phage display ScFv library against Mycobacterium tuberculosis].
OBJECTIVE
To construct a human phage display single-chain Fv (ScFv) antibody library against Mycobacterium tuberculosis (MTB), for specific ScFv antibody cloning.


METHODS
Total RNA was isolated from the lymphocytes of patients with positive serum antibody against MTB and reverse transcribed into cDNA. The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by PCR and then assembled into ScFv genes. The ScFv genes were ligated into phagemid pCANTAB5S. The human phage display ScFv library against MTB was constructed by transforming the recombinant phagemid into E. coli TG1 with the presence of helper phage M13K07.


RESULTS
The human phage display ScFv library containing 10(7) different clones was constructed successfully.


CONCLUSIONS
A phage display ScFv library against MTB has been constructed based on the variable region gene of immunoglobulin of the lymphocytes of TB patients and phagemid pCANTAB5S. The specific ScFv antibodies can be screened from this library.
Yang
Hua
H
Shanghai Key Laboratory of Tuberculosis, Pulmonary Hospital of Shanghai Affiliated Tongji University, Shanghai 200433, China.
Zhou
Xin-gang
XG
Shi
Li
L
Feng
Yong-hong
YH
Qin
Lian-hua
LH
Zheng
Rui-juan
RJ
Wang
Jie
J
Hu
Zhong-yi
ZY
D000911:Antibodies, Monoclonal; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000276:immunology* / Q000302:isolation & purification / Q000821:virology*; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000235:genetics*
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D057127:Single-Chain Antibodies