title: HPLC immunoaffinity purification of rabies virus glycoprotein using immobilized antipeptide antibodies.
abstract: It has been reported that the acetylcholine receptor may be used by the rabies virus to concentrate at sites in proximal to peripheral nerves. It has also been reported that the binding site for the receptor is located within the 190-203 region of the virus glycoprotein on the basis of its structural homology with the toxic center of snake neurotoxins, which are well known cholinergic ligands. We prepared monoclonal antibodies against the synthetic tetradecapeptide having the same sequence as the putative binding site of the rabies virus. One of three antibodies (clone 2PV 36-74) was able to recognize both the whole virus and its peplomeric glycoprotein and could bind acetylcholine. It was also able to inhibit the binding both of alpha-bungarotoxin and rabies virus glycoprotein to the acetylcholine receptor. We have covalently bound 2PV 36-74 to an HPLC affinity column and utilized it for specific purification of rabies virus glycoprotein. The immunoaffinity chromatographic method we describe is very sensitive and highly specific. Moreover this procedure does not denature the sample and is vary rapid and efficient.
authors.affiliation: CRISMA, Centro Didattico dell'Università, Siena, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Santucci
authors.forename: M
authors.initials: M
authors.lastname: Rustici
authors.forename: L
authors.initials: L
authors.lastname: Bracci
authors.forename: L
authors.initials: L
authors.lastname: Lozzi
authors.forename: P
authors.initials: P
authors.lastname: Soldani
authors.forename: P
authors.initials: P
authors.lastname: Neri
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D051379:Mice; D011820:Rabies virus / Q000032:analysis*; D014764:Viral Proteins / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D014764:Viral Proteins