Homogeneouzs Coenzyme-Immunoasssay for Quantitation of Insulin in Solution

Autor:	A. I. Gromov, A. M. Yegorov, Alexander P. Osipov, T.G. Mitrokhina, E. M. Gavrilova, Sorokina Nv
Rok vydání:	1982
Předmět:	biology Chemistry Insulin medicine.medical_treatment Biochemistry (medical) Clinical Biochemistry Biochemistry Binding constant Cofactor Analytical Chemistry Guinea pig chemistry.chemical_compound Electrochemistry medicine biology.protein NAD+ kinase Spectroscopy Acetamide Conjugate Carbodiimide
Zdroj:	Analytical Letters. 15:171-192
ISSN:	1532-236X 0003-2719
DOI:	10.1080/00032718208064374
Popis:	The Homogeneous Coenzyme-immunoassay for quantitation of insulin in a solution has been developed. The method relies on the comptitive binding of free and coenzyme (NAD)-labeled insulin by insulin antibodies (guinea pig anti-porcine insulin). NAD-insulin conjugates were obtained by a covalent binding of NAD-N6-[N-(6-aminohexy1)acetamide] and insulin in the presence of water-soluble carbodiimide. The NAD-insulin conjugaties were cycled in the coenzyme regeneration system Which included horese liver alcoholdehydrogenase (AIH) and its coupled substrates-cyclohexanol and P-nitroson-N, N-dimethlaniline (NIMA)- and the cycling rate was used to measure the conjugate concentrations. the cycling was inhibited when the lignad (insulin molecule) was bound to the insulin antibodey. The unbound insulin-NAD conjugate could be measure in the presence of the protein-bound cojugate. The Sensitivity of the proposed method of insulin determination in limited by the value of the antigen-sntibody binding constant and...
Databáze:	OpenAIRE
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