Title Broadly reactive monoclonal antibodies against beta-lactamases for immunodetection of bacterial resistance to antibiotics /
Authors Bielskė, Karolina ; Petraitytė-Burneikienė, Rasa ; Avižinienė, Aliona ; Dapkūnas, Justas ; Plikusienė, Ieva ; Juciutė, Silvija ; Stančiauskaitė, Miglė ; Žvirblienė, Aurelija ; Kučinskaitė-Kodzė, Indrė
DOI 10.1038/s41598-025-04603-2
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Is Part of Scientific reports.. Berlin : Springer Nature. 2025, vol. 15, iss. 1, art. no. 19094, p. [1-16].. ISSN 2045-2322. eISSN 2045-2322
Keywords [eng] AmpC β-lactamases ; Antibiotic resistance ; Class C β-lactamases ; Monoclonal antibodies ; β-lactamase detection
Abstract [eng] With antibiotic resistance reaching alarming levels globally, rapid detection of resistance determinants is crucial for administering appropriate antimicrobial therapies. This study aimed to develop monoclonal antibodies (MAbs) against bacterial β-lactamases, which are key enzymes in antibiotic resistance, for potential diagnostic use. To generate MAbs capable of recognising a broad range of β-lactamases in bacterial isolates, the bacteriophage vB_EcoS_NBD2 tail tube protein gp39-derived nanotubes, as a scaffold displaying a highly conserved 17-amino acid peptide of AmpC β-lactamases, were produced in yeast and used as an immunogen for generation of MAbs by hybridoma technology. Thirteen hybridoma clones producing peptide-specific MAbs were developed. To assess MAb reactivity with AmpC enzymes, recombinant DHA-1, PDC-195, ACT-14, CMY-34, and ADC-144 β-lactamases were generated. Eleven of thirteen MAbs demonstrated cross-reactivity with all tested β-lactamases in ELISA and Western blot. Immunoprecipitation and Western blot analyses confirmed MAb reactivity with natural CMY-34 in the Citrobacter portucalensis isolate. Epitope analysis revealed that most MAbs recognise a highly conserved epitope of 11 amino acids. The MAbs were comprehensively characterised using different immunoassays, total internal reflection ellipsometry and computational modelling. These novel MAbs, which recognise a wide range of AmpC enzymes, represent a promising tool for immunodetection of antibiotic resistance determinants.
Published Berlin : Springer Nature
Type Journal article
Language English
Publication date 2025
CC license CC license description