Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases

Cawez Frédéric, Paola Sandra Mercuri, Francisco Morales Yanez, Rita Maalouf, Marylène Vandevenne, Frederic Kerff, Virginie Guérin, View ORCID ProfileJacques Mainil, View ORCID ProfileDamien Thiry, Marc Saulmont, Alain Vanderplasschen, Pierre Lafaye, Gabriel Aymé, Pierre Bogaerts, Mireille Dumoulin, Moreno Galleni
doi: https://doi.org/10.1101/2022.07.01.498528
Cawez Frédéric
aInBioS, Center for Protein Engineering, Biological Macromolecules, Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Paola Sandra Mercuri
aInBioS, Center for Protein Engineering, Biological Macromolecules, Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Francisco Morales Yanez
bInBioS, Center for Protein Engineering, NEPTUNS (Nanobodies to Explore Protein Structure and Functions), Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Rita Maalouf
bInBioS, Center for Protein Engineering, NEPTUNS (Nanobodies to Explore Protein Structure and Functions), Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Marylène Vandevenne
cInBios, Center for Protein Engineering, ROBOTEIN@, Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Frederic Kerff
dInBioS, Center for Protein Engineering, Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Virginie Guérin
eBacteriology, FARAH and Faculty of Veterinary Medicine, Department of Infectious and Parasitic Diseases, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jacques Mainil
eBacteriology, FARAH and Faculty of Veterinary Medicine, Department of Infectious and Parasitic Diseases, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Jacques Mainil
Damien Thiry
eBacteriology, FARAH and Faculty of Veterinary Medicine, Department of Infectious and Parasitic Diseases, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Damien Thiry
Marc Saulmont
fRegional Animal Health and Identification Association (ARSIA), Ciney, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alain Vanderplasschen
gImmunology-Vaccinology, FARAH and Faculty of Veterinary Medicine, Department of Infectious and Parasitic Diseases, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Pierre Lafaye
hPlateforme d’Ingénierie des Anticorps, C2RT, Institut Pasteur, CNRS UMR 3528, Paris, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Gabriel Aymé
hPlateforme d’Ingénierie des Anticorps, C2RT, Institut Pasteur, CNRS UMR 3528, Paris, France
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Pierre Bogaerts
iNational reference center for antibiotic-resistant Gram-negative bacilli, Department of Clinical Microbiology, CHU UCL Namur, Yvoir, Namur, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mireille Dumoulin
bInBioS, Center for Protein Engineering, NEPTUNS (Nanobodies to Explore Protein Structure and Functions), Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Moreno Galleni
aInBioS, Center for Protein Engineering, Biological Macromolecules, Department of Life Sciences, University of Liège, Belgium
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: mgalleni@uliege.be
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

ABSTRACT

Soluble single-domain fragments derived from the unique variable region of camelid heavy-chain antibodies (VHHs) against enzymes may behave as potent inhibitors. The immunization of alpacas with the CMY-2 β-lactamase led to the isolation of three VHHs that specifically recognized and inhibited CMY-2. The structure of the complex VHH cAbCMY-2(254)/CMY-2 was determined by X-ray crystallography. We showed that the epitope is close to the active site and that the CDR3 of the VHH protrudes in the catalytic site. The β-lactamase inhibition was found to follow a mixed profile with a predominant non-competitive component. The three isolated VHHs recognized overlapping epitopes since they behaved as competitive binder. Our study identified a binding site that can be targeted by a new class of β-lactamase’s inhibitors designed with the help of a peptidomimetic approach. Furthermore, the use of mono or bivalent VHH and rabbit polyclonal anti-CMY-2 antibodies enable the development of the first generation of ELISA test for the detection of CMY-2 produced by resistant bacteria.

IMPORTANCE The still increasing antimicrobial resistance in human clinic or veterinary medicine is a major threat for modern chemotherapy. Beside the major caution in the use of current antibiotics, it is important to develop new classes of antibiotics. This work was focused on β-lactamases that are the enzymes involved in the hydrolysis of the major class of antibiotics, the β-lactam compounds. We selected camelid antibodies that inhibit CMY-2, a class C β-lactamase produced by bacteria isolated from the veterinary and human settings. We characterized the conformational epitope present in CMY-2 in order to create a new family of inhibitors based on the paratope of the antibody. Finally, we designed a primary version of a detection system based on an ELISA using VHH and polyclonal antibodies.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted July 03, 2022.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases
Cawez Frédéric, Paola Sandra Mercuri, Francisco Morales Yanez, Rita Maalouf, Marylène Vandevenne, Frederic Kerff, Virginie Guérin, Jacques Mainil, Damien Thiry, Marc Saulmont, Alain Vanderplasschen, Pierre Lafaye, Gabriel Aymé, Pierre Bogaerts, Mireille Dumoulin, Moreno Galleni
bioRxiv 2022.07.01.498528; doi: https://doi.org/10.1101/2022.07.01.498528
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases
Cawez Frédéric, Paola Sandra Mercuri, Francisco Morales Yanez, Rita Maalouf, Marylène Vandevenne, Frederic Kerff, Virginie Guérin, Jacques Mainil, Damien Thiry, Marc Saulmont, Alain Vanderplasschen, Pierre Lafaye, Gabriel Aymé, Pierre Bogaerts, Mireille Dumoulin, Moreno Galleni
bioRxiv 2022.07.01.498528; doi: https://doi.org/10.1101/2022.07.01.498528

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biochemistry
Subject Areas
All Articles
  • Animal Behavior and Cognition (3698)
  • Biochemistry (7809)
  • Bioengineering (5689)
  • Bioinformatics (21330)
  • Biophysics (10595)
  • Cancer Biology (8199)
  • Cell Biology (11961)
  • Clinical Trials (138)
  • Developmental Biology (6777)
  • Ecology (10419)
  • Epidemiology (2065)
  • Evolutionary Biology (13900)
  • Genetics (9726)
  • Genomics (13094)
  • Immunology (8164)
  • Microbiology (20058)
  • Molecular Biology (7871)
  • Neuroscience (43147)
  • Paleontology (321)
  • Pathology (1280)
  • Pharmacology and Toxicology (2264)
  • Physiology (3362)
  • Plant Biology (7246)
  • Scientific Communication and Education (1315)
  • Synthetic Biology (2010)
  • Systems Biology (5547)
  • Zoology (1132)