Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1986-11-20
1989-05-30
Kepplinger, Esther M.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 18, 435 19, 435 20, 435 21, 435 24, 435810, 436518, 436536, 436537, 436808, G01N 3353, G01N 33543, G01N 33536, C12Q 134
Patent
active
048350990
ABSTRACT:
A method for enzyme immunoassay for a ligand suspected to be present in a liquid sample includes signal amplification by use of at least two hydrolases and a blocked fluoroketone for one of the hydrolases. Ligand present in the liquid binds to an antiligand and a hydrolase-labeled tracer. The resulting bound fraction is separated and the hydrolase in the tracer removes the blocking group from the blocked fluoroketone. The fluoroketone activates or inhibits a second hydrolase which catalyzes the conversion of a substrate to a product. The presence or absence of the ligand in the liquid is indicated by a signal, such as a color change or a rate of color change, associated with the product. The invention includes a new class of hydrolase inhibitors and blocked fluoroketones and a kit of materials useful for performing the method of the invention.
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Gelb et al, Fluoro Ketone Inhibitors of Hydrolytic Enzymes, Biochemistry, vol. 24, No. 8, Apr. 9, 1985, pp. 1813-1817.
C. Walsh, Suicide Substrates: Mechanism Based Enzyme Inactivators Tetrahedron 38, 871 (1982).
Mize Patrick D.
O'Connell James P.
Becton Dickinson and Company
Brown Richard E.
Hoffer Florina B.
Kepplinger Esther M.
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