Dzantiev, B.B. Choi, Myung Ja Park, J. Choi, J. Romanenko, O.G. Zherdev, A.V. Eremin, S.A. Izumrudov, V.A. 2024-01-21T22:10:49Z 2024-01-21T22:10:49Z 2021-09-02 1994-01 0165-2478 https://pubs.kist.re.kr/handle/201004/145909 A new visual enzyme immunoassay (EIA) technique has been developed. Oppositely charged synthetic linear water-soluble polyelectrolytes (poly-N-ethyl-4-vinyl-pyridine as polycation and polymethacrylate as polyanion) were used as carriers for reagent immobilization. The ability of these molecules to form an insoluble complex was applied for the separation of bound and free components of the immunoassay reaction mixture. This approach was realized in methamphetamine visual EIA. In the first stage of the assay two specific reactions took place during incubation of the analytical reagents with the probe to be analyzed: (1) competition between methamphetamine and hapten conjugated with peroxidase for the interaction with specific antibodies and (2) interaction of these antibodies with the protein A-polymethacrylate conjugate. As a result of these reactions the (polyanion-protein A)-antibody-(hapten-peroxidase) complex was formed. Then the reaction mixture was filtered through an Ultrabind membrane (0.45 μm) with adsorbed poly-N-ethyl-4-vinylpyridine, and the immunological complexes were immobilized to the membrane by electrostatic interaction. The level of peroxidase binding on the membrane was measured by diaminobenzidene substrate. The system described was optimized to achieve both high rapidity (20 min) and an appropriate sensitivity (0.4 μg/ml) for methamphetamine assay. ？ 1994. English methamphetamine polyelectrolyte antibody production article enzyme immunoassay nonhuman priority journal rabbit Animal Electrolytes Haptens Horseradish Peroxidase Immune Sera Immunoenzyme Techniques Male Methamphetamine Polymethacrylic Acids Polyvinyls Pyridinium Compounds Rabbits Staphylococcal Protein A A new visual enzyme immunoassay of methamphetamine using linear water-soluble polyelectrolytes Article 10.1016/0165-2478(94)90134-1 1 Immunology Letters, v.41, no.2-3, pp.205 - 211 Immunology Letters 41 2-3 205 211 scopus 2-s2.0-0027937337 Article methamphetamine polyelectrolyte antibody production article enzyme immunoassay nonhuman priority journal rabbit Animal Electrolytes Haptens Horseradish Peroxidase Immune Sera Immunoenzyme Techniques Male Methamphetamine Polymethacrylic Acids Polyvinyls Pyridinium Compounds Rabbits Staphylococcal Protein A Enzyme immunoassay Membrane Methamphetamine Polyelectrolyte