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Molecular evolution of antibody cross-reactivity for two subtypes of type A botulinum neurotoxin

Department of Anesthesia and Pharmaceutical Chemistry, University of California, San Francisco Rm. 3C-38, San Francisco General Hospital, 1001 Potrero Ave., San Francisco, California 94110, USA.
Nature Biotechnology (Impact Factor: 39.08). 02/2007; 25(1):107-16. DOI: 10.1038/nbt1269
Source: PubMed

ABSTRACT 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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    • "Higher affinity antibodies were generated from the lead antibodies using molecular evolution or affinity maturation techniques [38] [39]. Once a selected scFv was affinity matured, it was converted to a full-length IgG as described previously [39]. The epitopes that the engineered antibodies bind have been defined [36,38,41–43] and are indicated in Supplemental Table 1. "
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    • "Higher affinity antibodies were generated from the lead antibodies using molecular evolution or affinity maturation techniques [38] [39]. Once a selected scFv was affinity matured, it was converted to a full-length IgG as described previously [39]. The epitopes that the engineered antibodies bind have been defined [36,38,41–43] and are indicated in Supplemental Table 1. "
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