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Production of recombinant OXA-23 carbapenemase: a target for developing antibody-based diagnostics against carbapenem-resistant Acinetobacter baumannii

Journal of Applied Biology and Biotechnology, ISSN: 2347-212X, Vol: 13, Issue: 1, Page: 291-298
2025
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Article Description

Carbapenems are typically the treatment of choice for drug-resistant Acinetobacter baumannii; however, the emergence of carbapenem-resistant strains has elevated this pathogen to the World Health Organization’s critical priority pathogen list. Since carbapenem resistance is frequently mediated by carbapenemases, particularly oxacillinases and metallo-beta-lactamases, antibody-based carbapenemase detection tests can be developed for rapid and affordable diagnosis of the infection. However, the development of such tests requires the availability of high-quality target proteins for generating specific antibodies, their characterization, and assay optimization. In this study, we reported a streamlined workflow to obtain a purified preparation of the tagless and functionally active recombinant OXA-23 enzyme, which is one of the major factors responsible for carbapenem resistance in Acinetobacter baumannii. The recombinant protein was expressed in the heterologous expression host Escherichia coli using auto-induction and purified using a single step of chromatography, followed by the removal of the affinity tag using TEV protease. The simple protocol reported here is expected to facilitate the production of other carbapenemases and similar proteins for the development of diagnostics and therapeutics to support the fight against antimicrobial-resistant bacteria.

Bibliographic Details

Abhishek Kasana; Brinda Bhardwaj; Vaishali Verma

Open Science Publishers LLP

Biochemistry, Genetics and Molecular Biology; Agricultural and Biological Sciences

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