1P4B

Three-Dimensional Structure Of a Single Chain Fv Fragment Complexed With The peptide GCN4(7P-14P).

1P4B の概要

• エントリーDOI: 10.2210/pdb1p4b/pdb
• 分子名称: Antibody Variable light chain, Antibody Variable heavy chain, GCN4(7P-14P) peptide, ... (4 entities in total)
• 機能のキーワード: fv antibody, antigen peptide binder, scfv, picomolar binder, immune system
• 由来する生物種: Mus musculus (house mouse); 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 28191.94

構造登録者

Zahnd, C.,Spinelli, S.,Luginbuhl, B.,Jermutus, L.,Amstutz, P.,Cambillau, C.,Pluckthun, A. (登録日: 2003-04-22, 公開日: 2004-05-04, 最終更新日: 2024-11-13)

主引用文献

Zahnd, C.,Spinelli, S.,Amstutz, P.,Cambillau, C.
Directed in Vitro Evolution and Crystallographic Analysis of a Peptide-binding Single Chain Antibody Fragment (scFv) with Low Picomolar Affinity.
J.Biol.Chem., 279:18870-18877, 2004

PubMed Abstract: 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 mutations, which do not make direct contact to the antigen, lead to a 500-fold affinity improvement over its potential germ line precursor. These results suggest that the affinity optimization of very high affinity binders is achieved by modulating existing interactions via subtle changes in the framework rather than by introducing new contacts. Off-rate selection in combination with ribosome display can evolve binders to the low picomolar affinity range even for peptide targets.

PubMed: 14754898
DOI: 10.1074/jbc.M309169200

実験手法

X-RAY DIFFRACTION (2.35 Å)