دورية أكاديمية
أعرض في EDS

Structure and specificity of an anti-chloramphenicol single domain antibody for detection of amphenicol residues.

التفاصيل البيبلوغرافية
العنوان: Structure and specificity of an anti-chloramphenicol single domain antibody for detection of amphenicol residues.
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.)
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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
الوصف
تدمد:1469-896X
DOI:10.1002/pro.4457