title: Purification of ornithine aminotransferase by immunoadsorption.
abstract: Ornithine aminotransferase was purified by conventional biochemical methods from rat kidney, rat liver, and human liver. Affinity-purified antibodies raised to the rat kidney enzyme were used to produce an immunoadsorbent enabling a one-step purification of ornithine aminotransferase to be made from crude human liver extracts. The harsh chemical conditions often required to desorb immunoadsorbents were avoided by isolating antibodies with low functional affinity and employing an electrophoretic desorption method which allowed the enzyme activity to be retained. The close structural similarity between human and rat ornithine aminotransferase was demonstrated by immunodiffusion reactions. It was therefore possible to purify the enzyme from human liver using immobilized antibodies raised against rat kidney ornithine aminotransferase. Furthermore, desorption was more readily achieved due to the lower affinity for the human enzyme.
authors.affiliation: Department of Life Sciences, Trent Polytechnic, Nottingham, United Kingdom.
authors.forename: J M
authors.initials: JM
authors.lastname: Leah
authors.forename: E E
authors.initials: EE
authors.lastname: Billett
authors.forename: T
authors.initials: T
authors.lastname: Palmer
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D004800:Enzymes, Immobilized; D006801:Humans; D007163:Immunosorbent Techniques; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D009953:Ornithine-Oxo-Acid Transaminase / Q000276:immunology / Q000302:isolation & purification*; D011485:Protein Binding; D051381:Rats; D013329:Structure-Activity Relationship; D000637:Transaminases / Q000302:isolation & purification*
chemical_list: D004800:Enzymes, Immobilized; D000637:Transaminases; D009953:Ornithine-Oxo-Acid Transaminase