2nyy

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==Overview==
==Overview==
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Broadening antibody specificity without compromising affinity should, facilitate detection and neutralization of toxin and viral subtypes. We, used yeast display and a co-selection strategy to increase, cross-reactivity of a single chain (sc) Fv antibody to botulinum, neurotoxin type A (BoNT/A). Starting with a scFv that binds the BoNT/A1, subtype with high affinity (136 pM) and the BoNT/A2 subtype with low, affinity (109 nM), we increased its affinity for BoNT/A2 1,250-fold, to 87, pM, while maintaining high-affinity binding to BoNT/A1 (115 pM). To find, the molecular basis for improved cross-reactivity, we determined the X-ray, co-crystal structures of wild-type and cross-reactive antibodies complexed, to BoNT/A1 at resolutions up to 2.6 A, and measured the thermodynamic, contribution of BoNT/A1 and A2 amino acids to wild-type and cross-reactive, antibody binding. The results show how an antibody can be engineered to, bind two different antigens despite structural differences in the, antigen-antibody interface and may provide a general strategy for tuning, antibody specificity and cross-reactivity.
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Broadening antibody specificity without compromising affinity should facilitate detection and neutralization of toxin and viral subtypes. We used yeast display and a co-selection strategy to increase cross-reactivity of a single chain (sc) Fv antibody to botulinum neurotoxin type A (BoNT/A). Starting with a scFv that binds the BoNT/A1 subtype with high affinity (136 pM) and the BoNT/A2 subtype with low affinity (109 nM), we increased its affinity for BoNT/A2 1,250-fold, to 87 pM, while maintaining high-affinity binding to BoNT/A1 (115 pM). To find the molecular basis for improved cross-reactivity, we determined the X-ray co-crystal structures of wild-type and cross-reactive antibodies complexed to BoNT/A1 at resolutions up to 2.6 A, and measured the thermodynamic contribution of BoNT/A1 and A2 amino acids to wild-type and cross-reactive antibody binding. The results show how an antibody can be engineered to bind two different antigens despite structural differences in the antigen-antibody interface and may provide a general strategy for tuning antibody specificity and cross-reactivity.
==About this Structure==
==About this Structure==
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[[Category: Clostridium botulinum]]
[[Category: Clostridium botulinum]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: Arndt, J.W.]]
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[[Category: Arndt, J W.]]
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[[Category: Stevens, R.C.]]
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[[Category: Stevens, R C.]]
[[Category: CA]]
[[Category: CA]]
[[Category: ZN]]
[[Category: ZN]]
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[[Category: protein antibody complex]]
[[Category: protein antibody complex]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri Feb 15 17:40:00 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:12:39 2008''

Revision as of 16:12, 21 February 2008


2nyy, resolution 2.61Å

Drag the structure with the mouse to rotate

Crystal structure of botulinum neurotoxin type A complexed with monoclonal antibody CR1

Overview

Broadening antibody specificity without compromising affinity should facilitate detection and neutralization of toxin and viral subtypes. We used yeast display and a co-selection strategy to increase cross-reactivity of a single chain (sc) Fv antibody to botulinum neurotoxin type A (BoNT/A). Starting with a scFv that binds the BoNT/A1 subtype with high affinity (136 pM) and the BoNT/A2 subtype with low affinity (109 nM), we increased its affinity for BoNT/A2 1,250-fold, to 87 pM, while maintaining high-affinity binding to BoNT/A1 (115 pM). To find the molecular basis for improved cross-reactivity, we determined the X-ray co-crystal structures of wild-type and cross-reactive antibodies complexed to BoNT/A1 at resolutions up to 2.6 A, and measured the thermodynamic contribution of BoNT/A1 and A2 amino acids to wild-type and cross-reactive antibody binding. The results show how an antibody can be engineered to bind two different antigens despite structural differences in the antigen-antibody interface and may provide a general strategy for tuning antibody specificity and cross-reactivity.

About this Structure

2NYY is a Single protein structure of sequence from Clostridium botulinum with and as ligands. Full crystallographic information is available from OCA.

Reference

Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin., Garcia-Rodriguez C, Levy R, Arndt JW, Forsyth CM, Razai A, Lou J, Geren I, Stevens RC, Marks JD, Nat Biotechnol. 2007 Jan;25(1):107-16. Epub 2006 Dec 17. PMID:17173035

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