| Download | - View final version: Isolation of TGF-β-neutralizing single-domain antibodies of predetermined epitope specificity using next-generation DNA sequencing (PDF, 693 KB)
- View supplementary information: Isolation of TGF-β-neutralizing single-domain antibodies of predetermined epitope specificity using next-generation DNA sequencing (DOCX, 202 KB)
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| DOI | Resolve DOI: https://doi.org/10.1093/protein/gzw043 |
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| Author | Search for: Henry, Kevin A.1; Search for: Hussack, Greg1; Search for: Collins, Cathy1; Search for: Zwaagstra, John C.1; Search for: Tanha, Jamshid1; Search for: MacKenzie, C. Roger1 |
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| Name affiliation | - National Research Council of Canada. Human Health Therapeutics
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| Format | Text, Article |
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| Subject | antibody; single-domain antibody; VHH; TGF-β; next-generation DNA sequencing; phage display |
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| Abstract | The 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. |
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| Publication date | 2016-09-08 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 23000759 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 646f2540-63b6-4b0e-a7ae-731f5c2a6082 |
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| Record created | 2016-09-14 |
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| Record modified | 2020-06-02 |
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