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Published on: October 15, 2016
[Screening of human anti-TNF-alpha scFv from large phage library]
Yu-xiao Wang1, Yuan-yuan Qiao, Li-jun Zhou
1Navy General Hospital, Beijing 100037, China.
Aim:
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.
Conclusion:
Human TNF-alpha scFv have been successfully obtained from large phage antibody library.
