NDLI: Phage display of human antibodies from a patient suffering from coeliac disease and selection of isotype-specific scFv against gliadin

Content Provider	PubMed Central
Author	Rhyner, Claudio Weichel, Michael Hübner, Philipp Achatz, Gernot Blaser, Kurt Crameri, Reto
Copyright Year	2003
Abstract	Coeliac disease (CD), a gastrointestinal illness characterized by intestinal malabsorption, results from gluten intolerance accompanied with immunological responses towards gliadin, an ethanol-soluble protein fraction of wheat and other cereals. The role of gliadin in eliciting immune responses in CD is still partly unclear; however, the occurrence of anti-gliadin in the sera of patients suffering from CD correlates well with clinical symptoms. In this work we report the construction of isotype-specific, phage-displayed scFv libraries from peripheral blood lymphocytes of a patient with CD and from a healthy control individual. VH and VL chains were amplified by reverse transcription–polymerase chain reaction (RT–PCR) using a set of oligonucleotides recognizing all human variable gene families. The three scFv libraries (IgA, IgG and IgM) were selectively enriched for gliadin-binding phage. After four rounds of affinity selection, polyclonal enrichment of gliadin-binding phage was observed in all libraries from the CD patient but in none from the healthy donor. Phagemid particles generated from single clones were demonstrated to be gliadin-specific, as shown by strongly positive enzyme-linked immunosorbent assay (ELISA) and BiaCore signals. The VH and VL chains from samples of these monoclonal isotype-specific phage were sequenced to identify the most common variable regions used by the immune system to elicit antibody responses against gliadin.
Related Links	http://dx.doi.org/10.1046/j.1365-2567.2003.01728.x
Ending Page	274
Page Count	6
Starting Page	269
File Format	PDF
ISSN	00192805
e-ISSN	13652567
Journal	Immunology
Issue Number	2
Volume Number	110
Language	English
Publisher	Blackwell Science Inc
Publisher Date	2003-10-01
Access Restriction	Open
Rights Holder	Blackwell Science Inc
Subject Keyword	Immunology Immunology and Allergy Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Immunology and Allergy Immunology

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