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A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells.

Posted by on in 2017
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Hundsberger H1Koppensteiner A1Hofmann E1Ripper D1Pflüger M1Stadlmann V2Klein CT1Kreiseder B1Katzlinger M3Eger A1Forster F2Missbichler A2Wiesner C1SLAS Discov. 2017 Mar 1:2472555217697435. doi: 10.1177/2472555217697435. [Epub ahead of print]

1
1 Department of Medical and Pharmaceutical Biotechnology, University of Applied Sciences Krems, Krems, Austria.
2
2 Sciotec Diagnostics Technologies GmbH, Tulln, Austria.
3
3 Molecular Devices, LLC, Wals-Siezenheim, Austria.

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.

KEYWORDS:

AlphaLISA; Caco-2 monolayer; ECIS; anti-gliadin IgY; celiac disease

PMID:
 
28277888
 
DOI:
 
10.1177/2472555217697435
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