Directed in vitro evolution and crystallographic analysis of a peptide-binding single chain antibody fragment (scFv) with low picomolar affinity.

@article{Zahnd2004DirectedIV,
  title={Directed in vitro evolution and crystallographic analysis of a peptide-binding single chain antibody fragment (scFv) with low picomolar affinity.},
  author={Christian Zahnd and Silvia Spinelli and B{\'e}atrice Luginb{\"u}hl and Patrick Amstutz and Christian Cambillau and Andreas Pl{\"u}ckthun},
  journal={The Journal of biological chemistry},
  year={2004},
  volume={279 18},
  pages={18870-7}
}
We generated a single chain Fv fragment of an antibody (scFv) with a binding affinity of about 5 pm to a short peptide by applying rigorous directed evolution. Starting from a high affinity peptide binder, originally obtained by ribosome display from a murine library, we generated libraries of mutants with error-prone PCR and DNA shuffling and applied off-rate selection by using ribosome display. Crystallographic analysis of the scFv in its antigen-bound and free state showed that only few… CONTINUE READING