title: Immunoaffinity purification of avermectin-binding proteins from the free-living nematode Caenorhabditis elegans and the fruitfly Drosophila melanogaster.
abstract: Avermectin-binding proteins from the free-living nematode worm Caenorhabditis elegans and from the fruitfly Drosophila melanogaster were purified to homogeneity via a three-step procedure. The binding proteins were covalently labelled using a radioactive photoaffinity probe and then partially purified on a Sephacryl S-300 gel-filtration column. The radiolabelled binding proteins were then purified by immunoaffinity chromatography using a monoclonal antibody to avermectin covalently attached to Protein A-Sepharose beads. Three affinity-labelled Drosophila proteins with molecular masses between 45 and 50 kDa were isolated in this way and then separated from each other by electroelution. This three-step protocol provides a rapid technique for receptor purification which may be of use in the purification of other binding proteins.
authors.affiliation: Department of Cellular Biochemistry and Physiology, Merck Research Laboratories, Rahway, NJ 07065.
authors.forename: S P
authors.initials: SP
authors.lastname: Rohrer
authors.forename: E B
authors.initials: EB
authors.lastname: Jacobson
authors.forename: E C
authors.initials: EC
authors.lastname: Hayes
authors.forename: E T
authors.initials: ET
authors.lastname: Birzin
authors.forename: J M
authors.initials: JM
authors.lastname: Schaeffer
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D017173:Caenorhabditis elegans / Q000737:chemistry*; D002352:Carrier Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004331:Drosophila melanogaster / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D007559:Ivermectin / Q000031:analogs & derivatives* / Q000378:metabolism; D011233:Precipitin Tests
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D007559:Ivermectin; C019264:avermectin