[Screening of human anti-TNF-alpha scFv from large phage library].
OBJECTIVE
To clone human anti-TNF-alpha scFv from large phage antibody library.


METHODS
Panning of large phage library against TNF-alpha was conducted to select specific antibodies. The antigen binding activity and DNA sequence were determined and analyzed by ELISA, restriction enzyme map.


RESULTS
After 4 rounds of panning, 62 positive clones were obtained and 27 clones had specific binding ability to TNF-alpha. DNA fingerprinting of the 27 clones showed 7 different stripes indicated 7 different positive clones. 5 of the 7 clones expressed soluble anti-TNF-alpha activity. DNA sequence analysis showed that the variable regions of these scFvs belonged to different subgroups.


CONCLUSIONS
Human TNF-alpha scFv have been successfully obtained from large phage antibody library.
Wang
Yu-xiao
YX
Navy General Hospital, Beijing 100037, China.
Qiao
Yuan-yuan
YY
Zhou
Li-jun
LJ
Wang
Yan
Y
D000906:Antibodies / Q000235:genetics / Q000276:immunology; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology*; D019151:Peptide Library; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology*
D000906:Antibodies; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D014409:Tumor Necrosis Factor-alpha