Enzyme coupled immunoassay of insulin using a novel coupling reagent.
A novel coupling reagent, meta-maleimidobenzoyl N-hydroxysuccinimide ester was synthesized. Using this reagent, insulin was conjugated very easily with beta-D-galactosidase [EC 3.2.1.23] in neutral, aqueous solution. No reduction of the enzyme activity was observed during the coupling procedure. The competitive bindings of the conjugate and insulin to anti-insulin serum were tested. The results indicated that the conjugate has enough immune reactivity for use in enzyme coupled immunoassay. Using this assay 20-800 pg of insulin were detectable.
D001667:Binding, Competitive; D005696:Galactosidases / Q000378:metabolism; D007118:Immunoassay; D007202:Indicators and Reagents; D007328:Insulin / Q000032:analysis*; D007330:Insulin Antibodies; D008301:Maleimides; D011485:Protein Binding; D013388:Succinimides
D007202:Indicators and Reagents; D007328:Insulin; D007330:Insulin Antibodies; D008301:Maleimides; D013388:Succinimides; D005696:Galactosidases