دورية أكاديمية
Structure and specificity of an anti-chloramphenicol single domain antibody for detection of amphenicol residues.
العنوان: | Structure and specificity of an anti-chloramphenicol single domain antibody for detection of amphenicol residues. |
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المؤلفون: | Swofford CA; Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Nordeen SA; Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Chen L; Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Desai MM; Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Chen J; Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Springs SL; Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Schwartz TU; Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Sinskey AJ; Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. |
المصدر: | Protein science : a publication of the Protein Society [Protein Sci] 2022 Nov; Vol. 31 (11), pp. e4457. |
نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, Non-U.S. Gov't |
اللغة: | English |
بيانات الدورية: | Publisher: Cold Spring Harbor Laboratory Press Country of Publication: United States NLM ID: 9211750 Publication Model: Print Cited Medium: Internet ISSN: 1469-896X (Electronic) Linking ISSN: 09618368 NLM ISO Abbreviation: Protein Sci Subsets: MEDLINE |
أسماء مطبوعة: | Publication: 2001- : Woodbury, NY : Cold Spring Harbor Laboratory Press Original Publication: New York, N.Y. : Cambridge University Press, c1992- |
مواضيع طبية MeSH: | Single-Domain Antibodies* , Camelids, New World*, Animals ; Chloramphenicol ; Reproducibility of Results ; Anti-Bacterial Agents ; Antibody Specificity |
مستخلص: | Antibiotics in aquaculture prevent bacterial infection of fish, but their misuse is a public health risk and contributes to the unintentional creation of multiresistant pathogens. Regulatory agencies cannot do the rigorous, expensive testing required to keep up with the volume of seafood shipments. Current rapid test kits for these drugs enable the increase in testing needed for adequate monitoring of food supply chains, but they lack a high degree of accuracy. To combat this, we set out to discover and engineer single-domain antibodies (VHHs) that bind to small molecule antibiotics, and that can be used in rapid test kits. The small size, solubility, and stability of VHHs are useful properties that can improve the reliability and shelf-life of test kits for these adulterants. Here, we report a novel anti-chloramphenicol VHH (Chl-VHH) with a disassociation constant of 57 nM. This was achieved by immunizing a llama against a chloramphenicol-keyhole limpet hemocyanin (KLH) conjugate and screening for high affinity binders through phage display. The crystal structure of the bound-VHH to chloramphenicol was key to identifying a mutation in the binding pocket that resulted in a 16-fold improvement in binding affinity. In addition, the structure provides new insights into VHH-hapten interactions that can guide future engineering of VHHs against additional targets. (© 2022 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.) |
References: | Elife. 2016 Jul 19;5:. (PMID: 27434673) Protein Sci. 2022 Nov;31(11):e4457. (PMID: 36153664) Anal Chim Acta. 2010 Jul 5;672(1-2):93-102. (PMID: 20579496) Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21. (PMID: 20057044) J Mol Biol. 2001 Aug 3;311(1):123-9. (PMID: 11469862) Anal Chem. 2017 Jun 20;89(12):6800-6806. (PMID: 28494149) J Immunol Methods. 2003 Oct 1;281(1-2):161-75. (PMID: 14580890) Methods Mol Biol. 2012;911:15-26. (PMID: 22886243) Protein Sci. 2011 Jul;20(7):1196-207. (PMID: 21557375) Anal Chem. 2008 Dec 15;80(24):9604-11. (PMID: 19072267) Sci Rep. 2019 Feb 12;9(1):1840. (PMID: 30755682) Environ Pollut. 2012 Mar;162:56-62. (PMID: 22243847) Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2015;32(11):1842-50. (PMID: 26359628) Elife. 2013 Sep 10;2:e01456. (PMID: 24040512) Anal Chem. 2014 Sep 2;86(17):8873-80. (PMID: 25079057) Lancet Infect Dis. 2016 Feb;16(2):161-8. (PMID: 26603172) Anal Bioanal Chem. 2016 Sep;408(22):5985-6002. (PMID: 27209591) Anal Chem. 2017 Jun 6;89(11):6248-6256. (PMID: 28460522) Food Chem Toxicol. 2017 Jun;104:3-13. (PMID: 27908699) Mol Immunol. 2008 Aug;45(14):3703-13. (PMID: 18632156) J Biol Chem. 2015 Apr 3;290(14):9087-100. (PMID: 25678711) Anal Chem. 2012 Jan 17;84(2):1165-71. (PMID: 22148739) Anal Chem. 2011 Sep 15;83(18):7213-20. (PMID: 21827167) Annu Rev Immunol. 2018 Apr 26;36:695-715. (PMID: 29490163) ACS Cent Sci. 2017 Feb 22;3(2):110-116. (PMID: 28280777) Biochemistry. 2000 Feb 15;39(6):1217-22. (PMID: 10684599) FEBS Lett. 2019 Jun;593(11):1248-1256. (PMID: 31049949) Nat Struct Mol Biol. 2018 Mar;25(3):289-296. (PMID: 29434346) Lancet Microbe. 2020 May;1(1):e34-e43. (PMID: 35538907) Anal Chem. 2014 Aug 19;86(16):8296-302. (PMID: 25068372) Acta Crystallogr D Struct Biol. 2019 Oct 1;75(Pt 10):861-877. (PMID: 31588918) J Mol Recognit. 2019 Jan;32(1):e2755. (PMID: 30033524) Anal Chem. 2006 Jul 1;78(13):4501-8. (PMID: 16808459) Mol Immunol. 2007 Mar;44(7):1680-90. (PMID: 17011035) |
معلومات مُعتمدة: | P30 GM124165 United States GM NIGMS NIH HHS |
فهرسة مساهمة: | Keywords: VHH; antibiotics; chloramphenicol; food adulteration; haptens; nanobodies; single-domain antibodies |
المشرفين على المادة: | 0 (Single-Domain Antibodies) 66974FR9Q1 (Chloramphenicol) 0 (Anti-Bacterial Agents) |
تواريخ الأحداث: | Date Created: 20220925 Date Completed: 20221101 Latest Revision: 20221109 |
رمز التحديث: | 20240628 |
مُعرف محوري في PubMed: | PMC9601811 |
DOI: | 10.1002/pro.4457 |
PMID: | 36153664 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1469-896X |
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DOI: | 10.1002/pro.4457 |