Selection of human single chain Fv antibody fragments binding and inhibiting Helicobacter pylori urease

Détails

ID Serval
serval:BIB_0F30801E947F
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Selection of human single chain Fv antibody fragments binding and inhibiting Helicobacter pylori urease
Périodique
Tumour Biology
Auteur(s)
Houimel  M., Corthesy-Theulaz  I., Fisch  I., Wong  C., Corthesy  B., Mach  J., Finnern  R.
ISSN
1010-4283 (Print)
Statut éditorial
Publié
Date de publication
02/2001
Volume
22
Numéro
1
Pages
36-44
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jan-Feb
Résumé
Single chain Fv antibody fragments (scFv) binding to purified Helicobacter pylori urease were selected from a nonimmune human antibody repertoire displayed on filamentous phage. After three rounds of screening on solid phase urease, 44 clones were found to bind the enzyme and four distinct scFv were identified by sequencing their heavy and light chain variable region genes (V(H) and V(L)). Two of the selected human scFv (scFv B4 and scFv D9) inhibited the activity of H. pylori urease with inhibitory constants (K(i)) of 7 and 2 microM, respectively. Their affinity (K(d)) for H. pylori urease as determined by surface plasmon resonance ranged from 17 to 42 nM. Both scFv were able to bind to urease present on the surface of living H. pylori organisms as demonstrated by flow cytometry analysis. The binding sites of scFv B4 and D9 were mapped by the use of two random hexapeptide libraries (X6 and CX6C) displayed on filamentous bacteriophage. The selected peptide sequences were shown to inhibit scFv binding to H. pylori urease and thus could be used in a vaccination strategy as epitopes mimicking (mimotopes) the region of urease recognized by these human scFv antibody fragments.
Mots-clé
Amino Acid Sequence Antibodies, Bacterial/*immunology/isolation & purification/metabolism/pharmacology Antibody Affinity Antibody Specificity Antigen-Antibody Reactions Bacterial Proteins/antagonists & inhibitors/*immunology/metabolism Bacterial Vaccines Drug Design Enzyme Inhibitors/immunology/isolation & purification Enzyme-Linked Immunosorbent Assay Epitopes/immunology Helicobacter pylori/enzymology/*immunology Humans Immunoglobulin Fragments/*immunology/isolation & purification/metabolism/pharmacology Molecular Mimicry Molecular Sequence Data Peptide Library Sequence Alignment Sequence Homology, Amino Acid Surface Plasmon Resonance Urease/antagonists & inhibitors/*immunology/metabolism
Pubmed
Web of science
Création de la notice
25/01/2008 14:53
Dernière modification de la notice
20/08/2019 12:35
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