title: Inhibition of biofilm formation by Camelid single-domain antibodies against the flagellum of Pseudomonas aeruginosa.
abstract: Pseudomonas aeruginosa is a leading cause of hospital-acquired infections in patients with compromised host defense mechanisms, including burn wound victims. In addition to its intrinsic resistance against most antibiotics, P. aeruginosa has the ability to form biofilms adhering to biotic or abiotic surfaces. These factors make treatment of P. aeruginosa infections complicated and demand new therapies and drugs. The flagellum of P. aeruginosa plays an important role in cell-cell and cell-surface interactions during the first stage of biofilm formation. In this study, we describe the selection of monoclonal anti-flagellin single-domain antibodies (VHHs) derived from the Camelid heavy-chain antibody repertoire of a llama immunized with P. aeruginosa antigens. The anti-flagellin VHHs could be produced efficiently in Saccharomyces cerevisiae, and surface plasmon resonance experiments demonstrated that they have apparent affinities in the nanomolar range. Functional screens showed that the anti-flagellin VHHs are capable of inhibiting P. aeruginosa from swimming and that they prevent biofilm formation in an in vitro assay. These data open doors for the development of novel methods for the prevention of P. aeruginosa-related infections.
authors.affiliation: BAC BV, Department R&D, J.H. Oortweg 21, 2333 CH Leiden, The Netherlands; Department of Molecular Microbiology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands. Electronic address: hendrik.adams@bac.nl.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Adams
authors.affiliation: BAC BV, Department R&D, J.H. Oortweg 21, 2333 CH Leiden, The Netherlands.
authors.forename: Wannie M
authors.initials: WM
authors.lastname: Horrevoets
authors.affiliation: BAC BV, Department R&D, J.H. Oortweg 21, 2333 CH Leiden, The Netherlands.
authors.forename: Simon M
authors.initials: SM
authors.lastname: Adema
authors.affiliation: BAC BV, Department R&D, J.H. Oortweg 21, 2333 CH Leiden, The Netherlands.
authors.forename: Hannah E V
authors.initials: HE
authors.lastname: Carr
authors.affiliation: Department of Molecular Microbiology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Richard E
authors.initials: RE
authors.lastname: van Woerden
authors.affiliation: Department of Molecular Microbiology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Margot
authors.initials: M
authors.lastname: Koster
authors.affiliation: Department of Molecular Microbiology, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: Tommassen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D002161:Camelids, New World; D005407:Flagella / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D005408:Flagellin / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa / Q000187:drug effects*; D016415:Sequence Alignment; D061905:Single-Domain Antibodies / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D061905:Single-Domain Antibodies; D005408:Flagellin
keywords: Biofilm; Camelid single-domain antibodies; Flagellum; Pseudomonas aeruginosa; VHH