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[Improving the affinity of an anti-TNF-alpha scFv by random mutation]
Bao-an Wang1, Xiao-sui Chen, Yu-xiao Wang
1Central Lab., Navy General Hospital, Beijing 100037, China.
Aim:
To improve the affinity of an anti-TNF-alpha scFv.
Methods:
A mutant phage antibody library derived from an anti-TNF-alpha scFv gene was generated by error-prone PCR. The mutated genes were then subjected to DNA shuffling. Mutants with improved affinity were selected by bio-panning. Affinity improvement of the selected mutants was verified by dot blot ELISA and thiocyanate elusion ELISA.
Results:
One mutant was obtained with relative affinity index (1.37 mol/L) higher than that of the parent scFv (0.48 mol/L).
Conclusion:
Error-prone PCR plus DNA shuffling is effective in improving the affinity of antibodies.
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