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- Title
Isolation and characterization of a VHH targeting the Acinetobacter baumannii cell surface protein CsuA/B.
- Authors
Lei, Eric K.; Ryan, Shannon; van Faassen, Henk; Foss, Mary; Robotham, Anna; Baltat, Isabel; Fulton, Kelly; Henry, Kevin A.; Chen, Wangxue; Hussack, Greg
- Abstract
Acinetobacter baumannii is a Gram-negative bacterial pathogen that exhibits high intrinsic resistance to antimicrobials, with treatment often requiring the use of last-resort antibiotics. Antibiotic-resistant strains have become increasingly prevalent, underscoring a need for new therapeutic interventions. The aim of this study was to use A. baumannii outer membrane vesicles as immunogens to generate single-domain antibodies (VHHs) against bacterial cell surface targets. Llama immunization with the outer membrane vesicle preparations from four A. baumannii strains (ATCC 19606, ATCC 17961, ATCC 17975, and LAC-4) elicited a strong heavy-chain IgG response, and VHHs were selected against cell surface and/or extracellular targets. For one VHH, OMV81, the target antigen was identified using a combination of gel electrophoresis, mass spectrometry, and binding studies. Using these techniques, OMV81 was shown to specifically recognize CsuA/B, a protein subunit of the Csu pilus, with an equilibrium dissociation constant of 17 nM. OMV81 specifically bound to intact A. baumannii cells, highlighting its potential use as a targeting agent. We anticipate the ability to generate antigen-specific antibodies against cell surface A. baumannii targets could provide tools for further study and treatment of this pathogen. Key points: •Llama immunization with bacterial OMV preparations for VHH generation •A. baumannii CsuA/B, a pilus subunit, identified by mass spectrometry as VHH target •High-affinity and specific VHH binding to CsuA/B and A. baumannii cells
- Subjects
ACINETOBACTER baumannii; EXTRACELLULAR vesicles; BACTERIAL cell surfaces; GRAM-negative bacteria; MASS spectrometry; BACTERIAL cell walls; PEPTIDE antibiotics; CELL separation
- Publication
Applied Microbiology & Biotechnology, 2023, Vol 107, Issue 14, p4567
- ISSN
0175-7598
- Publication type
Academic Journal
- DOI
10.1007/s00253-023-12594-1