1p4b
From Proteopedia
(New page: 200px<br /> <applet load="1p4b" size="450" color="white" frame="true" align="right" spinBox="true" caption="1p4b, resolution 2.35Å" /> '''Three-Dimensional S...) |
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| - | [[Image:1p4b.gif|left|200px]]<br /> | + | [[Image:1p4b.gif|left|200px]]<br /><applet load="1p4b" size="350" color="white" frame="true" align="right" spinBox="true" |
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caption="1p4b, resolution 2.35Å" /> | caption="1p4b, resolution 2.35Å" /> | ||
'''Three-Dimensional Structure Of a Single Chain Fv Fragment Complexed With The peptide GCN4(7P-14P).'''<br /> | '''Three-Dimensional Structure Of a Single Chain Fv Fragment Complexed With The peptide GCN4(7P-14P).'''<br /> | ||
==Overview== | ==Overview== | ||
| - | We generated a single chain Fv fragment of an antibody (scFv) with a | + | 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. |
==About this Structure== | ==About this Structure== | ||
| - | 1P4B is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Mus_musculus Mus musculus]. Full crystallographic information is available from [http:// | + | 1P4B is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Mus_musculus Mus musculus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1P4B OCA]. |
==Reference== | ==Reference== | ||
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[[Category: scfv]] | [[Category: scfv]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:24:56 2008'' |
Revision as of 12:24, 21 February 2008
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Three-Dimensional Structure Of a Single Chain Fv Fragment Complexed With The peptide GCN4(7P-14P).
Overview
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.
About this Structure
1P4B 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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