Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1987-04-09
1989-08-15
Kepplinger, Esther M.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435808, 436517, 436807, 436533, 436534, 436518, 356427, G01N 3353, G01N 33557, G01N 33546, G01N 33543
Patent
active
048574546
ABSTRACT:
A method is described for kinetic measurement of enzyme activity bound to a solid matrix which improves both the sensitivity and speed of one immunoassay method. The immunoassay typically consists of reaction of the analyte with two specific antibodies, one fixed to the surface of a polymeric bead or wall of a test tube, the other added in solution and labeled by covalent coupling to an enzyme. By reaction between analyte and both antibodies, the enzyme-labeled antibody becomes fixed to the surface in a quantity proportional to the quantity of the analyte. After washing sufficiently to remove unreacted enzyme-labeled antibody, fixed enzyme activity is measured by incubation with a substrate and measurement of the rate of the reaction catalyzed. Fixation of the enzyme causes the reaction products to be localized near the surface. To measure the concentration of reactant or product repeatedly during the reaction, the solution must be mixed before each measurement, which can interfere with the measurement. In the prior art, the reaction is stopped after incubation and the product measured once. The method and apparatus disclosed here provides stirring and measurement away from the surface, and thus permits repeated measurement during the reaction. This kinetic assay can be performed more rapidly and sensitively than assays based on a single measurement.
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Myrtle J., et al., "Creatine Kinase MB Isoenzyme Quantitation in Serum by Simultaneous Immunoenzymetric Assay", Clin. Chem., 29, 1232, (1983).
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Freundlich Lawrence F.
Karmen Arthur
Murthy Vadiraja V.
Albert Einstein College of Medicine of Yeshiva University a Division of
Deck Randall E.
Kepplinger Esther M.
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