Application of Multilayer Analytical Film to ELISA of Vanilmandelic Acid

✍ Scribed by M. Yoshioka; C. Aso; T. Hiraoka; M. Kitajima; M. Kuroda; M. Sugi; H. Parvez

• Book ID: 102615229
• Publisher: Elsevier Science
• Year: 1994
• Tongue: English
• Weight: 339 KB
• Volume: 49
• Category: Article
• ISSN: 0026-265X
• DOI: 10.1006/mchj.1994.1033

✦ Synopsis

The urinary level of vanilmandelic acid (VMA) is one marker for diagnosis of neurblastoma in neonates. A large number of samples are needed for mass screening. In order to make a convenient and sophisticated assay, we examined a method based on dry chemistry by using a monoclonal antibody to VMA. The antibody adsorbed on the surface of the well of a microplate was equilibrated with VMA labeled with glucose oxidase (GOD-VMA). Free VMA in a sample competes with GOD-VMA for the binding sites of the antibody. An aliquot containing the excess GOD-VMA was spotted onto a multilayer film consisting of a spreading layer containing glucose and a reaction layer containing peroxidase and a leucodye. Hydrogen peroxide produced with GOD-VMA in the spreading layer penetrated the reaction layer and produced a blue dye. The amount of dye formed was measured by a reflection photometer. From a standard curve, VMA was measurable from 0.1 to (50 \mu \mathrm{g} / \mathrm{ml}). The values obtained by the present method were correlated with those by high-performance liquid chromatography and enzyme-linked immunosorbent assay. ( 1994 Academic Press, Inc.