A method for assay for an unknown enzyme suspected to be present in a liquid includes signal amplification by use of a second enzyme and a blocked modulator for the second enzyme. Unknown enzyme in the liquid removes a blocking group from the blocked modulator. The resulting modulator activates or inhibits the second enzyme which catalyzes an indicator reaction in which a substrate is converted to a product. The presence or absence of the unknown enzyme in the liquid is indicated by a signal, such as a color change or a rate of color change, associated with the indicator reaction. The concentration of the enzyme in the sample may be determined by the measurement of the signal. The invention includes a kit of materials useful for performing the method of the invention. The method involves the hydrolyzing of a blocked fluoroketone inhibitor to facilitate the analytical method.
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 enzymes and a blocked modulator for one of the enzymes. Ligand present in the liquid binds to an antiligand and an enzyme-labeled tracer. The resulting bound fraction is separated and the enzyme in the tracer removes the blocking group from the blocked modulator. The modulator activates or inhibits a second enzyme 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 enzyme inhibitors and blocked inhibitors and a kit of materials useful for performing the method of the invention.
Signal enhancement in immunoassay by modulation of enzymatic catalysis
申请人:Becton, Dickinson and Company
公开号:US04835099A1
公开(公告)日:1989-05-30
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.