Enzyme-linked sandwich immunoassay of ornithine delta-aminotransferase from rat liver using antibody-coupled glass rods as solid phase.
A macromolecular antigen, ornithine delta-aminotransferase [EC 2.6.1.13] from rat liver (OAT) was assayed by the sandwich procedure using rabbit (anti-OAT) Fab'-beta-D-galactosidase complex and rabbit (anti-OAT) IgG-coupled glass rods as a solid phase. The Fab' fragments of the rabbit (anti-OAT) IgG were conjugated with beta-D-galactosidase [EC 3.2.1.23] from Escherichia coli using N, N'-o-phenylenedimaleimide. The rabbit (anti-OAT) IgG was coupled to the aminoalkylsilyl glass rods using glutaraldehyde. The rabbit (anti-OAT) IgG-coupled glass rods were incubated with OAT and then with the rabbit (anti-OAT) Fab'-beta-D-galactosidase complex. The amount of OAT was determined from the activity of beta-D-galactosidase bound to the glass rods. A minimum of 0.03 fmoles of OAT could be determined by this method and use of the glass rods gave greater reproducibility, and was more sensitive and simpler than use of Sepharose 4B.
D000818:Animals; D000906:Antibodies; D005069:Evaluation Studies as Topic; D005898:Glass; D007124:Immunoenzyme Techniques; D008099:Liver / Q000201:enzymology*; D009953:Ornithine-Oxo-Acid Transaminase / Q000032:analysis* / Q000276:immunology; D011485:Protein Binding; D011817:Rabbits / Q000276:immunology; D051381:Rats; D000637:Transaminases / Q000032:analysis*
D000906:Antibodies; D000637:Transaminases; D009953:Ornithine-Oxo-Acid Transaminase