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

Harald Hundsberger; Anita Koppensteiner; Elisabeth Hofmann; Doris Ripper; Maren Pflüger; Valerie Stadlmann; Christian Theodor Klein; B. Kreiseder; M. Katzlinger; Andreas Eger; Florian Forster; A. Missbichler; Christoph Wiesner

doi:10.1177/2472555217697435

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

Harald Hundsberger, Anita Koppensteiner, Elisabeth Hofmann, Doris Ripper, Maren Pflüger, Valerie Stadlmann, Christian Theodor Klein, B. Kreiseder, M. Katzlinger, Andreas Eger, Florian Forster, A. Missbichler, Christoph Wiesner

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	1035-1043
Number of pages	9
Journal	SLAS Discovery
Volume	22
Issue number	8
DOIs	https://doi.org/10.1177/2472555217697435
Publication status	Published - 8 Sept 2017

Keywords

gliadin antibody
immunoglobulin Y antibody
interleukin 8
neutralizing antibody
recombinant tumor necrosis factor
transcription factor RelA
unclassified drug
gliadin
immunoglobulin
immunoglobulin enhancer binding protein
immunoglobulin Y
transcription factor
antibody screening
antibody specificity
Article
binding affinity
Caco-2 cell line
cell function
cell membrane permeability
cell survival
cell viability assay
controlled study
down regulation
electric cell substrate impedance sensing
enzyme linked immunosorbent assay
high throughput screening
human
human cell
impedance
intestine epithelium
permeability barrier
protein expression
protein phosphorylation
Western blotting
cell communication
cytology
epithelium cell
immunology
inflammation
intestine
isolation and purification
metabolism
pathology
procedures
Antibodies
Neutralizing
Caco-2 Cells
Cell Communication
Cell Survival
Down-Regulation
Epithelial Cells
Gliadin
High-Throughput Screening Assays
Humans
Immunoglobulins
Inflammation
Interleukin-8
Intestines
NF-kappa B
Transcription Factors
AlphaLISA
Caco-2 monolayer
celiac disease
anti-gliadin IgY
ECIS

IMC Research Focuses

Medical biotechnology

ÖFOS 2012 - Austrian Fields of Study

304005 Medical biotechnology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1177/2472555217697435

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Hundsberger, H., Koppensteiner, A., Hofmann, E., Ripper, D., Pflüger, M., Stadlmann, V., Klein, C. T., Kreiseder, B., Katzlinger, M., Eger, A., Forster, F., Missbichler, A., & Wiesner, C. (2017). A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells: SLAS Discovery. SLAS Discovery, 22(8), 1035-1043. https://doi.org/10.1177/2472555217697435

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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.",

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author = "Harald Hundsberger and Anita Koppensteiner and Elisabeth Hofmann and Doris Ripper and Maren Pfl{\"u}ger and Valerie Stadlmann and Klein, {Christian Theodor} and B. Kreiseder and M. Katzlinger and Andreas Eger and Florian Forster and A. Missbichler and Christoph Wiesner",

note = "Funding Information: The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Austrian FFG (Forschungs F{\"o}rderungs Gesellschaft) Project COIN, Nr. 286415 to Christoph Wiesner and Project BRIDGE, Nr. 2647553 to Harald Hundsberger. Publisher Copyright: {\textcopyright} 2017, {\textcopyright} 2017 Society for Laboratory Automation and Screening.",

year = "2017",

month = sep,

day = "8",

doi = "10.1177/2472555217697435",

language = "English",

volume = "22",

pages = "1035--1043",

journal = "SLAS Discovery",

issn = "2472-5552",

publisher = "Society for Laboratory Automation and Screening",

number = "8",

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Hundsberger, H , Koppensteiner, A, Hofmann, E, Ripper, D, Pflüger, M, Stadlmann, V, Klein, CT, Kreiseder, B, Katzlinger, M, Eger, A, Forster, F, Missbichler, A & Wiesner, C 2017, 'A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells: SLAS Discovery', SLAS Discovery, vol. 22, no. 8, pp. 1035-1043. https://doi.org/10.1177/2472555217697435

TY - JOUR

T1 - A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells

T2 - SLAS Discovery

AU - Hundsberger, Harald

AU - Koppensteiner, Anita

AU - Hofmann, Elisabeth

AU - Ripper, Doris

AU - Pflüger, Maren

AU - Stadlmann, Valerie

AU - Klein, Christian Theodor

AU - Kreiseder, B.

AU - Katzlinger, M.

AU - Eger, Andreas

AU - Forster, Florian

AU - Missbichler, A.

AU - Wiesner, Christoph

N1 - Funding Information: The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Austrian FFG (Forschungs Förderungs Gesellschaft) Project COIN, Nr. 286415 to Christoph Wiesner and Project BRIDGE, Nr. 2647553 to Harald Hundsberger. Publisher Copyright: © 2017, © 2017 Society for Laboratory Automation and Screening.

PY - 2017/9/8

Y1 - 2017/9/8

N2 - 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.

AB - 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.

KW - gliadin antibody

KW - immunoglobulin Y antibody

KW - interleukin 8

KW - neutralizing antibody

KW - recombinant tumor necrosis factor

KW - transcription factor RelA

KW - unclassified drug

KW - gliadin

KW - immunoglobulin

KW - immunoglobulin enhancer binding protein

KW - immunoglobulin Y

KW - transcription factor

KW - antibody screening

KW - antibody specificity

KW - Article

KW - binding affinity

KW - Caco-2 cell line

KW - cell function

KW - cell membrane permeability

KW - cell survival

KW - cell viability assay

KW - controlled study

KW - down regulation

KW - electric cell substrate impedance sensing

KW - enzyme linked immunosorbent assay

KW - high throughput screening

KW - human

KW - human cell

KW - impedance

KW - intestine epithelium

KW - permeability barrier

KW - protein expression

KW - protein phosphorylation

KW - Western blotting

KW - cell communication

KW - cytology

KW - epithelium cell

KW - immunology

KW - inflammation

KW - intestine

KW - isolation and purification

KW - metabolism

KW - pathology

KW - procedures

KW - Antibodies

KW - Neutralizing

KW - Caco-2 Cells

KW - Cell Communication

KW - Cell Survival

KW - Down-Regulation

KW - Epithelial Cells

KW - Gliadin

KW - High-Throughput Screening Assays

KW - Humans

KW - Immunoglobulins

KW - Inflammation

KW - Interleukin-8

KW - Intestines

KW - NF-kappa B

KW - Transcription Factors

KW - AlphaLISA

KW - Caco-2 monolayer

KW - celiac disease

KW - anti-gliadin IgY

KW - ECIS

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U2 - 10.1177/2472555217697435

DO - 10.1177/2472555217697435

M3 - Article

SN - 2472-5552

VL - 22

SP - 1035

EP - 1043

JO - SLAS Discovery

JF - SLAS Discovery

IS - 8

ER -

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

Abstract

Keywords

IMC Research Focuses

ÖFOS 2012 - Austrian Fields of Study

UN SDGs

Access to Document

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