1p4i: Difference between revisions

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[[Image:1p4i.gif|left|200px]]
{{Seed}}
[[Image:1p4i.png|left|200px]]


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{{STRUCTURE_1p4i|  PDB=1p4i  |  SCENE=  }}  
{{STRUCTURE_1p4i|  PDB=1p4i  |  SCENE=  }}  


'''Crystal Structure of scFv against peptide GCN4'''
===Crystal Structure of scFv against peptide GCN4===




==Overview==
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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.
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{{ABSTRACT_PUBMED_14754898}}


==About this Structure==
==About this Structure==
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[[Category: Picomolar binder]]
[[Category: Picomolar binder]]
[[Category: Scfv]]
[[Category: Scfv]]
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Revision as of 10:26, 29 July 2008

File:1p4i.png

Template:STRUCTURE 1p4i

Crystal Structure of scFv against peptide GCN4

Template:ABSTRACT PUBMED 14754898

About this Structure

1P4I is a Protein complex structure of sequences from Mus musculus. Full crystallographic information is available from OCA.

Reference

Directed in vitro evolution and crystallographic analysis of a peptide-binding single chain antibody fragment (scFv) with low picomolar affinity., Zahnd C, Spinelli S, Luginbuhl B, Amstutz P, Cambillau C, Pluckthun A, J Biol Chem. 2004 Apr 30;279(18):18870-7. Epub 2004 Jan 30. PMID:14754898

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