| Download | - View accepted manuscript: Protease-resistant single-domain antibodies inhibit Campylobacter jejuni motility (PDF, 1.8 MiB)
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| DOI | Resolve DOI: https://doi.org/10.1093/protein/gzu011 |
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| Author | Search for: Hussack, Greg1; Search for: Riazi, Ali; Search for: Ryan, Shannon1; Search for: van Faassen, Henk1; Search for: Mackenzie, Roger1; Search for: Tanha, Jamshid1; Search for: Arbabi-Ghahroudi, Mehdri1 |
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| Affiliation | - National Research Council of Canada. Human Health Therapeutics
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| Format | Text, Article |
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| Subject | C. jejuni; motility; protease resistance; protein stability; VHH |
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| Abstract | Camelid heavy-chain antibody variable domains (VHHs) are emerging as potential antimicrobial reagents. We have engineered a previously isolated VHH (FlagV1M), which binds Campylobacter jejuni flagella, for greater thermal and proteolytic stability. Mutants of FlagV1M were obtained from an error-prone polymerase chain reaction library that was panned in the presence of gastrointestinal (GI) proteases. Additional FlagV1M mutants were obtained through disulfide-bond engineering. Each approach produced V HHs with enhanced thermal stability and protease resistance. When the beneficial mutations from both approaches were combined, a hyperstabilized VHH was created with superior stability. The hyperstabilized V HH bound C. jejuni flagella with wild-type affinity and was capable of potently inhibiting C. jejuni motility in assays performed after sequential digestion with three major GI proteases, demonstrating the remarkable stability imparted to the VHH by combining our engineering approaches. |
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| Publication date | 2014-04-16 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NRC number | NRC-HHT-53217 |
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| NPARC number | 21272210 |
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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 | 8c8f81ab-1aab-424c-8d30-13add7ed0b72 |
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| Record created | 2014-07-23 |
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| Record modified | 2020-06-04 |
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