Identification of gliadin-binding peptides by phage display

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Chen, TingsuGothenburg University,Göteborgs universitet,Institutionen för cell- och molekylärbiologi,Department of Cell and Molecular Biology,University of Gothenburg (författare)

Identification of gliadin-binding peptides by phage display

Artikel/kapitelEngelska2011

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2011-02-17
Springer Science and Business Media LLC,2011
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LIBRIS-ID:oai:gup.ub.gu.se/137927
https://gup.ub.gu.se/publication/137927URI
https://doi.org/10.1186/1472-6750-11-16DOI
https://research.chalmers.se/publication/137927URI

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Språk:engelska

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BACKGROUND: Coeliac disease (CD) is a common and complex disorder of the small intestine caused by intolerance to wheat gluten and related edible cereals like barley and rye. Peptides originating from incomplete gliadin digestion activate the lamina propria infiltrating T cells to release proinflammatory cytokines, which in turn cause profound tissue remodelling of the small intestinal wall. There is no cure for CD except refraining from consuming gluten-containing products. RESULTS: Phage from a random oligomer display library were enriched by repeated pannings against immobilised gliadin proteins. Phage from the final panning round were plated, individual plaques picked, incubated with host bacteria, amplified to a population size of 1011 to 1012 and purified. DNA was isolated from 1000 purified phage populations and the region covering the 36 bp oligonucleotide insert from which the displayed peptides were translated, was sequenced. Altogether more than 150 different peptide-encoding sequences were identified, many of which were repeatedly isolated under various experimental conditions. Amplified phage populations, each expressing a single peptide, were tested first in pools and then one by one for their ability to inhibit binding of human anti-gliadin antibodies in ELISA assays. These experiments showed that several of the different peptide-expressing phage tested inhibited the interaction between gliadin and anti-gliadin antibodies. Finally, four different peptide-encoding sequences were selected for further analysis, and the corresponding 12-mer peptides were synthesised in vitro. By ELISA assays it was demonstrated that several of the peptides inhibited the interaction between gliadin molecules and serum anti-gliadin antibodies. Moreover, ELISA competition experiments as well as dot-blot and western blot revealed that the different peptides interacted with different molecular sites of gliadin. CONCLUSIONS: We believe that several of the isolated and characterised gliadin-binding peptides described here could provide valuable tools for researchers in the field of CD by facilitating studies on localisation and uptake of various gliadin peptides in the small intestine. In future work, the potential of these peptides to detoxify gluten will be investigated.

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MEDICIN OCH HÄLSOVETENSKAP Medicinsk bioteknologi hsv//swe
MEDICAL AND HEALTH SCIENCES Medical Biotechnology hsv//eng

Biuppslag (personer, institutioner, konferenser, titlar ...)

Hoffmann, Karolina,1980Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)hoffmann (författare)
Östman, Sofia M,1974Gothenburg University,Göteborgs universitet,Institutionen för biomedicin, avdelningen för infektionssjukdomar,Institute of Biomedicine, Department of Infectious Medicine,University of Gothenburg(Swepub:gu)xostms (författare)
Sandberg, Ann-Sofie,1951Chalmers tekniska högskola,Chalmers University of Technology(Swepub:cth)annsandb (författare)
Olsson, Olof,1950Gothenburg University,Göteborgs universitet,Institutionen för växt- och miljövetenskaper,Department of Plant and Environmental Sciences,University of Gothenburg(Swepub:gu)xoolov (författare)
Göteborgs universitetInstitutionen för cell- och molekylärbiologi (creator_code:org_t)

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Ingår i:BMC Biotechnology: Springer Science and Business Media LLC111472-6750

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Av författaren/redakt...: Chen, Tingsu; Hoffmann, Karoli ...; Östman, Sofia M, ...; Sandberg, Ann-So ...; Olsson, Olof, 19 ...

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