title: Selection and characterization of recombinant dromedary antiovalbumin antibody fragments that do not cross-react with ovalbumin-related protein X: use for immunoaffinity.
abstract: Ovalbumin-related protein X (OVAX) and ovalbumin are two very close ovalbumin-related serpins. As primary data on OVAX remain recent, information about possible cross-reaction of available antiovalbumin antibodies with OVAX is still missing. Using labeled purified OVAX and dot ligand blotting, we identified 49 recombinant dromedary antiovalbumin single domain antibody (sdAb) fragments that were unable to bind OVAX. Discrimination between OVAX and ovalbumin was confirmed for two of the corresponding sdAb fragments by surface plasmon resonance and Western ligand blotting (WLB) characterizations. Furthermore, they were covalently linked to Sepharose and used as an affinity matrix for ovalbumin depletion. At least 90% of the original ovalbumin was eliminated from the allantoic fluid of 14 day old chicken embryo in one step. These sdAb fragments, which bind ovalbumin with nanomolar affinity, should also contribute to a better characterization of ovalbumin preparations.
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, F-37380 Nouzilly, France. Gilles.Bruneau@tours.inra.fr
authors.forename: Gilles
authors.initials: G
authors.lastname: Bruneau
authors.forename: Abdelhaq
authors.initials: A
authors.lastname: Anouassi
authors.forename: Sophie
authors.initials: S
authors.lastname: Réhault-Godbert
authors.forename: Sylvie
authors.initials: S
authors.lastname: Canépa
authors.forename: Michel R
authors.initials: MR
authors.lastname: Blanc
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D015153:Blotting, Western; D002162:Camelus / Q000276:immunology*; D002642:Chick Embryo; D003429:Cross Reactions / Q000235:genetics / Q000276:immunology*; D007457:Iodine Radioisotopes / Q000032:analysis; D010047:Ovalbumin / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D015843:Serpins / Q000276:immunology; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins; D015843:Serpins; D010047:Ovalbumin