title: A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells.
abstract: Celiac disease (CD) is a chronic inflammatory condition caused by the ingestion of gliadin-containing food in genetically susceptible individuals. Undigested peptides of gliadin exert various effects, including increased intestinal permeability and inflammation in the small intestine. Although many therapeutic approaches are in development, a gluten-free diet is the only effective treatment for CD. Affecting at least 1% of the population in industrialized countries, it is important to generate therapeutic options against CD. Here, we describe the establishment of a high-throughput screening (HTS) platform based on AlphaLISA and electrical cell-substrate impedance sensing (ECIS) technology for the identification of anti-inflammatory and barrier-protective compounds in human enterocytes after pepsin-trypsin-digested gliadin (PT-gliadin) treatment. Our results show that the combination of these HTS technologies enables fast, reliable, simple, and label-free screening of IgY antibodies against PT-gliadin. Using this platform, we have identified a new chicken anti-PT-gliadin IgY antibody as a potential anti-CD agent.
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Harald
authors.initials: H
authors.lastname: Hundsberger
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Anita
authors.initials: A
authors.lastname: Koppensteiner
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Hofmann
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Doris
authors.initials: D
authors.lastname: Ripper
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Maren
authors.initials: M
authors.lastname: Pflüger
authors.affiliation: 2 Sciotec Diagnostics Technologies GmbH, Tulln, Austria.
authors.forename: Valerie
authors.initials: V
authors.lastname: Stadlmann
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Christian Theodor
authors.initials: CT
authors.lastname: Klein
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Birgit
authors.initials: B
authors.lastname: Kreiseder
authors.affiliation: 3 Molecular Devices, LLC, Wals-Siezenheim, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Katzlinger
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Andreas
authors.initials: A
authors.lastname: Eger
authors.affiliation: 2 Sciotec Diagnostics Technologies GmbH, Tulln, Austria.
authors.forename: Florian
authors.initials: F
authors.lastname: Forster
authors.affiliation: 2 Sciotec Diagnostics Technologies GmbH, Tulln, Austria.
authors.forename: Albert
authors.initials: A
authors.lastname: Missbichler
authors.affiliation: 1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Wiesner
mesh_terms: D057134:Antibodies, Neutralizing / Q000032:analysis*; D018938:Caco-2 Cells; D002450:Cell Communication; D002470:Cell Survival; D015536:Down-Regulation; D004847:Epithelial Cells / Q000166:cytology*; D005903:Gliadin / Q000276:immunology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007136:Immunoglobulins / Q000302:isolation & purification; D007249:Inflammation / Q000473:pathology; D016209:Interleukin-8 / Q000378:metabolism; D007422:Intestines / Q000166:cytology*; D016328:NF-kappa B / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D057134:Antibodies, Neutralizing; C016512:IgY; D007136:Immunoglobulins; D016209:Interleukin-8; D016328:NF-kappa B; D014157:Transcription Factors; D005903:Gliadin
keywords: AlphaLISA; Caco-2 monolayer; ECIS; anti-gliadin IgY; celiac disease