2.50
Hdl Handle:
http://hdl.handle.net/10033/621034
Title:
Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity.
Authors:
Kuhn, Philipp; Thiem, Stefanie; Steinert, Michael; Purvis, Duncan; Lugmayr, Veronika; Treutlein, Ulrich; Plobner, Lutz; Leiser, Robert-Matthias; Hust, Michael; Dübel, Stefan
Abstract:
Legionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella, alveolar macrophages can be infected, which may lead to a life-threatening pneumonia called Legionnaires' disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology. In this study, an antibody against L. pneumophila has been generated from the naïve human HAL antibody libraries by phage-display for the first time. The panning was performed on whole bacterial cells in order to select antibodies that bind specifically to the cell surface of untreated Legionella. The bacterial cell wall component lipopolysaccharide (LPS) was identified as the target structure. Specific binding to the important pathogenic L. pneumophila strains Corby, Philadelphia-1 and Knoxville was observed, while no binding was detected to seven members of the families Enterobacteriaceae, Pseudomonadaceae or Clostridiaceae. Production of this antibody in the recombinant scFv-Fc format using either a murine or a human Fc part allowed the set-up of a sandwich-ELISA for detection of Legionella cells. The scFv-Fc construct proved to be very stable, even when stored for several weeks at elevated temperatures. A sensitivity limit of 4,000 cells was achieved. The scFv-Fc antibody pair was integrated on a biosensor, demonstrating the specific and fast detection of L. pneumophila on a portable device. With this system, 10,000 Legionella cells were detected within 35 min. Combined with a water filtration/concentration system, this antibody may be developed into a promising reagent for rapid on-site Legionella monitoring.
Affiliation:
Helmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
Human Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity. 2017, 26 (1):29-38 Hum Antibodies
Journal:
Human antibodies
Issue Date:
19-Jul-2017
URI:
http://hdl.handle.net/10033/621034
DOI:
10.3233/HAB-170318
PubMed ID:
28582852
Type:
Article
Language:
en
ISSN:
1875-869X
Appears in Collections:
publications of the scientific administration (GFW)

Full metadata record

DC FieldValue Language
dc.contributor.authorKuhn, Philippen
dc.contributor.authorThiem, Stefanieen
dc.contributor.authorSteinert, Michaelen
dc.contributor.authorPurvis, Duncanen
dc.contributor.authorLugmayr, Veronikaen
dc.contributor.authorTreutlein, Ulrichen
dc.contributor.authorPlobner, Lutzen
dc.contributor.authorLeiser, Robert-Matthiasen
dc.contributor.authorHust, Michaelen
dc.contributor.authorDübel, Stefanen
dc.date.accessioned2017-08-02T12:02:25Z-
dc.date.available2017-08-02T12:02:25Z-
dc.date.issued2017-07-19-
dc.identifier.citationHuman Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity. 2017, 26 (1):29-38 Hum Antibodiesen
dc.identifier.issn1875-869X-
dc.identifier.pmid28582852-
dc.identifier.doi10.3233/HAB-170318-
dc.identifier.urihttp://hdl.handle.net/10033/621034-
dc.description.abstractLegionella are Gram-negative bacteria that are ubiquitously present in natural and man-made water reservoirs. When humans inhale aerosolized water contaminated with Legionella, alveolar macrophages can be infected, which may lead to a life-threatening pneumonia called Legionnaires' disease. Due to the universal distribution of Legionella in water and their potential threat to human health, the Legionella concentration in water for human use must be strictly monitored, which is difficult since the standard detection still relies on lengthy cultivation and analysis of bacterial morphology. In this study, an antibody against L. pneumophila has been generated from the naïve human HAL antibody libraries by phage-display for the first time. The panning was performed on whole bacterial cells in order to select antibodies that bind specifically to the cell surface of untreated Legionella. The bacterial cell wall component lipopolysaccharide (LPS) was identified as the target structure. Specific binding to the important pathogenic L. pneumophila strains Corby, Philadelphia-1 and Knoxville was observed, while no binding was detected to seven members of the families Enterobacteriaceae, Pseudomonadaceae or Clostridiaceae. Production of this antibody in the recombinant scFv-Fc format using either a murine or a human Fc part allowed the set-up of a sandwich-ELISA for detection of Legionella cells. The scFv-Fc construct proved to be very stable, even when stored for several weeks at elevated temperatures. A sensitivity limit of 4,000 cells was achieved. The scFv-Fc antibody pair was integrated on a biosensor, demonstrating the specific and fast detection of L. pneumophila on a portable device. With this system, 10,000 Legionella cells were detected within 35 min. Combined with a water filtration/concentration system, this antibody may be developed into a promising reagent for rapid on-site Legionella monitoring.en
dc.language.isoenen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleHuman Anti-Lipopolysaccharid (LPS) antibodies against Legionella with high species specificity.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for infection research, Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalHuman antibodiesen
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