Isolation of TGF-β-neutralizing single-domain antibodies of predetermined epitope specificity using next-generation DNA sequencing

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DOIResolve DOI: http://doi.org/10.1093/protein/gzw043
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TypeArticle
Journal titleProtein Engineering, Design and Selection
ISSN1741-0126
1741-0134
Pages15
Subjectantibody; single-domain antibody; VHH; TGF-β; next-generation DNA sequencing; phage display
AbstractThe epitope specificity of therapeutic antibodies is often critical to their efficacy and mode of action. Here, we report the isolation of single-domain antibodies (sdAbs) against a pre-specified epitope of TGF-β3: namely, the site of interaction between the cytokine and its cell-surface type II receptor. By panning a phage-displayed immune llama VhH library against TGF-β3 using competitive elution with soluble dimeric type II receptor ectodomain in tandem with next-generation DNA sequencing, we identified several sdAbs that competed with the receptor for TGF-β3 binding and neutralized TGF-β3 in in vitro cellular assays. In contrast, all other sdAbs identified using conventional panning approaches (i.e., without regard to epitope specificity) did not target the site of receptor:cytokine interaction. We expect this strategy to be generally applicable for identifying epitope-specific sdAbs when binding reagents directed against the epitope of interest are available. The sdAbs identified here are of potential interest as cancer immunotherapeutics.
Publication date
PublisherOxford University Press
LanguageEnglish
AffiliationAutomotive and Surface Transportation; National Research Council Canada; Human Health Therapeutics
Peer reviewedYes
NPARC number23000759
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Record identifier646f2540-63b6-4b0e-a7ae-731f5c2a6082
Record created2016-09-14
Record modified2016-09-14
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