Kit for conducting an assay to detect a substance using enzymati

Details Kit for conducting an assay to detect a substance using enzymati Kit for conducting an assay to detect a substance using enzymati

1996-12-16

1999-02-02

Elliott, George C.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

252700, 549510, C12N 900, C07D32100

Patent

active

058663894

ABSTRACT:
A kit including a dioxetane and an enzyme reactive therewith is useful in an assay method in which a member of a specific binding pair is detected by means of an optically detectable reaction, the improvement wherein the optically detectable reaction includes the reaction, with the enzyme, the dioxetane having the formula ##STR1## where T is a cycloalkyl or polycycloalkyl group bonded to the 4-membered ring portion of the dioxetane by a spiro linkage; Y is a fluorescent chromophore; X is H, alkyl, aryl, aralkyl, alkaryl, heteroalkyl, heteroaryl, cycloalkyl, cycloheteroalkyl, or enzyme-cleavable group; and Z is H or an enzyme-cleavable group, provided that at least one of X or Z must be an enzyme-cleavable group, so that the enzyme cleaves the enzyme-cleavable group from the dioxetane to form a negatively charged substituent bonded to the dioxetane, the negatively charged substituent causing the dioxetane to decompose to form a luminescent substance that includes group Y of said dioxetane.

REFERENCES:
patent: 4857652 (1989-08-01), Schaap
patent: 4962192 (1990-10-01), Schaap
patent: 4978614 (1990-12-01), Bronstein
patent: 5330900 (1994-07-01), Bronstein et al.
patent: 5538847 (1996-07-01), Bronstein et al.
patent: 5582980 (1996-12-01), Bronstein et al.
patent: 5605795 (1997-02-01), Bronstein et al.
Affidavit of Charles M. Zepp, Submitted as attachment to paper 13 in parent application 06/889,823, Apr. 1987.

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