Construction and expression of specificity-improved single-chain variable fragments against the bioactive naphthoquinone, plumbagin.

@article{Sakamoto2009ConstructionAE,
  title={Construction and expression of specificity-improved single-chain variable fragments against the bioactive naphthoquinone, plumbagin.},
  author={Seiichi Sakamoto and Futoshi Taura and Waraporn Putalun and Benyakan Pongkitwitoon and Ryota Tsuchihashi and Satoshi Morimoto and Junei Kinjo and Yukihiro Shoyama and Hiroyuki Tanaka},
  journal={Biological & pharmaceutical bulletin},
  year={2009},
  volume={32 3},
  pages={434-9}
}
We constructed a single-chain variable fragment (scFv) antibody against plumbagin (PL) with improved specific binding to PL. Variable heavy- and light-chain genes were cloned directly from the cDNA of hybridoma cell line 3A3 and assembled using the splice-overlap extension polymerase chain reaction (SOE-PCR) with specific primers including flexible peptide (Gly(4)Ser)(3) linker primers. The constructed scFv gene was ligated into the pET28a expression vector and transformed into Escherichia coli… CONTINUE READING

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