Development of Nanobodies as Theranostic Agents against CMY-2-Like Class C β-Lactamases

Frédéric Cawez; Paola Sandra Mercuri; Francisco Javier Morales-Yãnez; 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

doi:10.1128/aac.01499-22

Development of Nanobodies as Theranostic Agents against CMY-2-Like Class C β-Lactamases

Frédéric Cawez
, Paola Sandra Mercuri
, Francisco Javier Morales-Yãnez
, 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^*

^*Corresponding author for this work

Chemical Engineering

Research output: Contribution to journal › Article › peer-review

Abstract

Three soluble single-domain fragments derived from the unique variable region of camelid heavy-chain antibodies (VHHs) against the CMY-2 β-lactamase behaved as inhibitors. The structure of the complex VHH cAb_CMY-2(254)/CMY-2 showed that the epitope is close to the active site and that the CDR3 of the VHH protrudes into the catalytic site. The β-lactamase inhibition pattern followed a mixed profile with a predominant noncompetitive component. The three isolated VHHs recognized overlapping epitopes since they behaved as competitive binders. Our study identified a binding site that can be targeted by a new class of β-lactamase inhibitors designed on the sequence of the paratope. Furthermore, the use of mono- or bivalent VHH and rabbit polyclonal anti-CMY-2 antibodies enables the development of the first generation of enzyme-linked immunosorbent assay (ELISA) for the detection of CMY-2 produced by CMY-2-expressing bacteria, irrespective of resistotype.

Original language	English
Article number	e0149922
Number of pages	19
Journal	Antimicrobial Agents and Chemotherapy
Volume	67
Issue number	4
Early online date	9 Mar 2023
DOIs	https://doi.org/10.1128/aac.01499-22
Publication status	Published - Apr 2023

Keywords

Animals
Rabbits
Single-Domain Antibodies
Precision Medicine
beta-Lactamases/genetics
beta-Lactamase Inhibitors
Penicillins
Antibodies
Epitopes

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10.1128/aac.01499-22

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Cawez, F., Mercuri, P. S., Morales-Yãnez, F. J., Maalouf, R., Vandevenne, M., Kerff, F., Guérin, V., Mainil, J., Thiry, D., Saulmont, M., Vanderplasschen, A., Lafaye, P., Aymé, G., Bogaerts, P., Dumoulin, M., & Galleni, M. (2023). Development of Nanobodies as Theranostic Agents against CMY-2-Like Class C β-Lactamases. Antimicrobial Agents and Chemotherapy, 67(4), Article e0149922. https://doi.org/10.1128/aac.01499-22

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abstract = "Three soluble single-domain fragments derived from the unique variable region of camelid heavy-chain antibodies (VHHs) against the CMY-2 β-lactamase behaved as inhibitors. The structure of the complex VHH cAbCMY-2(254)/CMY-2 showed that the epitope is close to the active site and that the CDR3 of the VHH protrudes into the catalytic site. The β-lactamase inhibition pattern followed a mixed profile with a predominant noncompetitive component. The three isolated VHHs recognized overlapping epitopes since they behaved as competitive binders. Our study identified a binding site that can be targeted by a new class of β-lactamase inhibitors designed on the sequence of the paratope. Furthermore, the use of mono- or bivalent VHH and rabbit polyclonal anti-CMY-2 antibodies enables the development of the first generation of enzyme-linked immunosorbent assay (ELISA) for the detection of CMY-2 produced by CMY-2-expressing bacteria, irrespective of resistotype.",

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author = "Fr{\'e}d{\'e}ric Cawez and Mercuri, \{Paola Sandra\} and Morales-Y{\~a}nez, \{Francisco Javier\} and Rita Maalouf and Maryl{\`e}ne Vandevenne and Frederic Kerff and Virginie Gu{\'e}rin and Jacques Mainil and Damien Thiry and Marc Saulmont and Alain Vanderplasschen and Pierre Lafaye and Gabriel Aym{\'e} and Pierre Bogaerts and Mireille Dumoulin and Moreno Galleni",

year = "2023",

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doi = "10.1128/aac.01499-22",

language = "English",

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journal = "Antimicrobial Agents and Chemotherapy",

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Cawez, F, Mercuri, PS, Morales-Yãnez, FJ, Maalouf, R, Vandevenne, M, Kerff, F, Guérin, V, Mainil, J, Thiry, D, Saulmont, M, Vanderplasschen, A, Lafaye, P, Aymé, G, Bogaerts, P, Dumoulin, M & Galleni, M 2023, 'Development of Nanobodies as Theranostic Agents against CMY-2-Like Class C β-Lactamases', Antimicrobial Agents and Chemotherapy, vol. 67, no. 4, e0149922. https://doi.org/10.1128/aac.01499-22

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T1 - Development of Nanobodies as Theranostic Agents against CMY-2-Like Class C β-Lactamases

AU - Cawez, Frédéric

AU - Mercuri, Paola Sandra

AU - Morales-Yãnez, Francisco Javier

AU - Maalouf, Rita

AU - Vandevenne, Marylène

AU - Kerff, Frederic

AU - Guérin, Virginie

AU - Mainil, Jacques

AU - Thiry, Damien

AU - Saulmont, Marc

AU - Vanderplasschen, Alain

AU - Lafaye, Pierre

AU - Aymé, Gabriel

AU - Bogaerts, Pierre

AU - Dumoulin, Mireille

AU - Galleni, Moreno

PY - 2023/4

Y1 - 2023/4

N2 - Three soluble single-domain fragments derived from the unique variable region of camelid heavy-chain antibodies (VHHs) against the CMY-2 β-lactamase behaved as inhibitors. The structure of the complex VHH cAbCMY-2(254)/CMY-2 showed that the epitope is close to the active site and that the CDR3 of the VHH protrudes into the catalytic site. The β-lactamase inhibition pattern followed a mixed profile with a predominant noncompetitive component. The three isolated VHHs recognized overlapping epitopes since they behaved as competitive binders. Our study identified a binding site that can be targeted by a new class of β-lactamase inhibitors designed on the sequence of the paratope. Furthermore, the use of mono- or bivalent VHH and rabbit polyclonal anti-CMY-2 antibodies enables the development of the first generation of enzyme-linked immunosorbent assay (ELISA) for the detection of CMY-2 produced by CMY-2-expressing bacteria, irrespective of resistotype.

AB - Three soluble single-domain fragments derived from the unique variable region of camelid heavy-chain antibodies (VHHs) against the CMY-2 β-lactamase behaved as inhibitors. The structure of the complex VHH cAbCMY-2(254)/CMY-2 showed that the epitope is close to the active site and that the CDR3 of the VHH protrudes into the catalytic site. The β-lactamase inhibition pattern followed a mixed profile with a predominant noncompetitive component. The three isolated VHHs recognized overlapping epitopes since they behaved as competitive binders. Our study identified a binding site that can be targeted by a new class of β-lactamase inhibitors designed on the sequence of the paratope. Furthermore, the use of mono- or bivalent VHH and rabbit polyclonal anti-CMY-2 antibodies enables the development of the first generation of enzyme-linked immunosorbent assay (ELISA) for the detection of CMY-2 produced by CMY-2-expressing bacteria, irrespective of resistotype.

KW - Animals

KW - Rabbits

KW - Single-Domain Antibodies

KW - Precision Medicine

KW - beta-Lactamases/genetics

KW - beta-Lactamase Inhibitors

KW - Penicillins

KW - Antibodies

KW - Epitopes

U2 - 10.1128/aac.01499-22

DO - 10.1128/aac.01499-22

M3 - Article

C2 - 36892280

SN - 0066-4804

VL - 67

JO - Antimicrobial Agents and Chemotherapy

JF - Antimicrobial Agents and Chemotherapy

IS - 4

M1 - e0149922

ER -

Development of Nanobodies as Theranostic Agents against CMY-2-Like Class C β-Lactamases

Abstract

Keywords

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