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

SLAS Discov. 2017 Sep;22(8):1035-1043. doi: 10.1177/2472555217697435. Epub 2017 Mar 9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Neutralizing / analysis*
  • Caco-2 Cells
  • Cell Communication
  • Cell Survival
  • Down-Regulation
  • Epithelial Cells / cytology*
  • Gliadin / immunology*
  • High-Throughput Screening Assays / methods*
  • Humans
  • Immunoglobulins / isolation & purification
  • Inflammation / pathology
  • Interleukin-8 / metabolism
  • Intestines / cytology*
  • NF-kappa B / metabolism
  • Transcription Factors / metabolism

Substances

  • Antibodies, Neutralizing
  • IgY
  • Immunoglobulins
  • Interleukin-8
  • NF-kappa B
  • Transcription Factors
  • Gliadin