Abstract: A method and apparatus for displaying data presents results from a single assay across multiple wells and plates on a single screen in a format that preserves the spatial relationship between the wells on a plate and the sequential relationship between plates. Also provided to the user is the ability to display these results from multiple wells in a color coded format corresponding to a binary (e.g. good/bad) or quadripartite (e.g. good/marginally good/marginally bad/bad) division. Also displayed are plate-wide statistics, such as signal-to-noise ratio and variability, for multiple plates on a single screen so that problem plates can be readily identified. The ability for a user to view a summary of performance on a well-by-well basis across all compounds/plates is also provided to aid the user in detecting general trends, including the possible existence of mechanical errors. Finally, the user is also provided with the ability to select or deselect plates and assays for review in the above formats.

Abstract: A competitive assay for the concentration of a cyclic nucleotide such as cyclic adenosine monophosphate combines, in a reaction chamber provided with a capture antibody, an antibody for the cAMP or other cyclic nucleotide, the sample to be assayed and a conjugate of cAMP and alkaline phosphatase. The mixture is incubated and washed, and after washing, a 1,2-dioxetane which is caused to decompose when contacted by the alkaline phosphatase of said conjugate is added. The light emission caused by decomposition is measured, with the strength of the signal being inversely related to the concentration of the cyclic nucleotide. Optionally, an enhancement agent in the form of a polymeric onium salt may be added, to further enhance the light emission.

Abstract: A method of measuring the activity of at least two reporter gene products in an aliquot of a sample extract is disclosed. The activities of a first and second reporter enzyme are quantified by measuring the light signal produced by degradation of a first substrate by the first reporter enzyme and the light signal produced by the degradation of a second substrate by a second reporter enzyme. Both quantifications are sequentially performed on the same aliquot of sample extract.

Abstract: A method of measuring the activity of at least two reporter gene products in an aliquot of a sample extract is disclosed. The activities of a first and second reporter enzyme are quantified by measuring the light signal produced by degradation of a first substrate by the first reporter enzyme and the light signal produced by the degradation of a second substrate by a second reporter enzyme. Both quantifications are sequentially performed on the same aliquot of sample extract.

Abstract: The present invention discloses multiple enzyme assays which measure the activity of at least one endogenous enzyme in a single aliquot and a method of measuring the activity of multiple enzymes in an aliquot of a cell extract, wherein at least one of the enzymes is an endogenous enzyme. In one embodiment of the invention the activity of a first enzyme is quantified by measuring the light signal produced by degradation of a first enzyme substrate by the first enzyme and the activity of the second enzyme is quantified by measuring the light signal produced by the degradation of a second substrate. In the method of the present invention, both quantifications are performed on the same aliquot of cell extract. Different embodiments of the present invention provide for the detection of more than one endogenous enzyme and for the detection of at least one reporter enzyme and at least one endogenous enzyme. The present invention also discloses kits for detecting the activity of multiple enzymes.

Abstract: The present invention discloses chemiluminescent 1,2-dioxetane substrates capable of reacting with a neuraminidase to release optically detectable energy. These 1,2-dioxetanes have the general formula: wherein Z is and the variables are selected so as to induce decomposition of said dioxetane accompanied by chemiluminescence where Z is cleaved by neuraminidase present.

Abstract: A luminescence detecting apparatus and method for analyzing luminescent samples is disclosed. Luminescent samples are placed in a plurality of sample wells in a tray, and the tray is placed in a visible-light impervious chamber containing a charge coupled device camera. The samples may be injected in the wells, and the samples may be injected with buffers and reagents, by an injector. In the chamber, light from the luminescent samples pass through a collimator, a Fresnel field lens, a filter, and a camera lens, whereupon a focused image is created by the optics on the charge-coupled device (CCD) camera. The use of a Fresnel field lens, in combination with a collimator and filter, reduces crosstalk between samples below the level attainable by the prior art. Preferred embodiments of the luminescence detecting apparatus and method disclosed include central processing control of all operations, multiple wavelength filter wheel, and robot handling of samples and reagents.

Abstract: A kit for detecting a first substance in a sample including a stabilized 1,2-dioxetane bearing an enzyme-labile substituent, which is destabilized and caused to decompose by contacting the 1,2-dioxetane with an enzyme under conditions which cause the enzyme to cleave the enzyme-labile substituent from the dioxetane, thereby yielding a negatively charged oxygen anion bonded to the 1,2-dioxetane, which causes the 1,2-dioxetane to decompose without input from an external excitation energy source, the decomposition being accompanied by chemiluminescence; and a second component selected from the group consisting of a specific affinity substance (e.g., an antigen, an antibody or a nucleic acid probe) and an enzyme which destabilizes said 1,2-dioxetane.

Abstract: Probes labeled with 1,2-dioxetane precursors can be employed in a variety of assays. The probes may be nucleic acid, peptide nucleic acid, proteins including enzyme, antibody or antigen, steroid, carbohydrate, drug or non-drug hapten. The probe is provided with a 1,2-dioxetane precursor bound thereto, generally either covalently, or a strong ligand bond. The dioxetane precursor moiety is converted to a bound 1,2-dioxetane by exposure to singlet oxygen. These dioxetane (labels) either spontaneously decompose, or are induced to decompose by an appropriate trigger to release light. The trigger may be a change in pH temperature, or an agent which removes a protective group. Assay formats in which these 1,2-dioxetane labeled probes and referents may be used to include hybridization assays, immuno assays, gel-based assays and Capillary Zone Electrophoresis.

Abstract: Chemiluminescent 1,2-dioxetane compounds capable of producing light energy when decomposed, substantially stable at room temperature, represented by the formulas I or II:

Abstract: Spiroadamantyl dioxetanes bearing an alkoxy substituent, and an aromatic substituent of phenyl or naphthyl on the dioxetane ring can be activated to chemiluminesce if the aromatic substituent bears a moiety designated OX, wherein the X is cleaved by an enzyme with which the dioxetane is permitted to come in contact with. The T½ kinetics of the chemiluminescent reaction, as well as the signal intensity, or quantum yield of the chemiluminescent reaction, can be altered by selection of an electron-withdrawing or an electron-donating group Z, at positions on the aromatic substituent other than those adjacent the point of attachment to the dioxetane. Signal strength can further be enhanced by recognized chemiluminescent enhancers.

Abstract: A kit for detecting a first substance in a sample including a stabilized 1,2-dioxetane bearing an enzyme-labile substituent, which is destabilized and caused to decompose by contacting the 1,2-dioxetane with an enzyme under conditions which cause the enzyme to cleave the enzyme-labile group from the dioxetane, thereby yielding a negatively charged oxygen anion bonded to the 1,2-dioxetane, which causes the 1,2-dioxetane to decompose without input from an external excitation energy source, the decomposition being accompanied by chemiluminescence; and a second component selected from the group consisting of a specific affinity substance (e.g., an antigen, an antibody or a nucleic acid probe) and an enzyme which destabilizes said 1,2-dioxetane.

Abstract: A chemiluminescent assays for the determination of the presence or amount of a biopolymer in bound assays using 1,2-dioxetanes in connection with AttoPhos™ as chemiluminescent substrates for enzyme-labeled targets or probes is provided. Further disclosed is a kit for conducting a bioassay for the presence or concentration of a biopolymer comprising a) an enzyme complex; b) a 1,2-dioxetane; and c) AttoPhos™.

Abstract: Heterogenous and homogenous assays are provided for the detection of protease inhibitory activity in a sample or target compound, taking advantage of the chemiluminescent characteristics of 1,2-dioxetanes. In the heterogenous assay, a peptide bearing a cleavage site for the protease of interest is provided with a first member of a first ligand binding pair at one end, and a first member of a second ligand binding pair at the other end. The other member of the first ligand binding pair is attached to a surface, which binds the peptide, or protease substrate, to the surface. The peptide substrate is combined with the protease and target compound or sample. Substrate cleavage, if not inhibited, is allowed to occur, and any unbound cleaved fragments are removed. An enzyme complexed with the second member of the second ligand binding pair is added, and allowed to bind to any of the (uncleaved) first member of the second ligand binding pair remaining.

Abstract: An assay for the presence of luciferase in a biological sample offers heightened sensitivity, signal intensity and persistence. The assay is sensitive down to 50 fg luciferase. The biological sample is combined with essential ingredients luciferin, ADP, myokinase and Mg++. The myokinase converts ADP to ATP, necessary for the luciferase reaction, and AMP, which retards the reaction kinetics. The resulting assay exhibits a persistent glow emission which makes it adaptable to automation.

Abstract: Xanthan esters and acridans are substrates for horseradish peroxidase. These stable, enzymatically cleavable chemiluminescent esters are substrates for horseradish peroxidase which, together with peroxide is among the extensively used enzyme in enzyme-linked detection methods, including immunoassays, oligonucleotide detection and nucleic acid hybridization. The novel compounds are used, together with peroxide, alkali and the peroxidase, to indicate the presence and/or concentration of target compounds. The assays may be enhanced by the use of polymeric quaternary onium enhancement compounds or similar compounds selected to enhance the chemiluminescence emitted.

Abstract: Spiroadamantyl dioxetanes bearing an alkoxy substituent, and an aromatic substituent of phenyl or naphthyl on the dioxetane ring can be activated to chemiluminesce if the aromatic substituent bears a moiety designated OX, wherein the X is cleaved by an enzyme with which the dioxetane is permitted to come in contact with. The T.sub.1/2 kinetics of the chemiluminescent reaction, as well as the signal intensity, or quantum yield of the chemiluminescent reaction, can be altered by selection of an electron-withdrawing or an electron-donating group Z, at positions on the aromatic substituent other than those adjacent the point of attachment to the dioxetane. Signal strength can further be enhanced by recognized chemiluminescent enhancers.

Abstract: Dioxetanes that can be triggered by bases are provided which can be used to detect bases in organic solvents, aqueous preparations and detect gaseous bases in the atmosphere. The dioxetanes can be used to detect the release of bases from various labels, as a means to detect the presence of a base released by physical or natural processes, to calibrate light measuring apparatus and to determine the amount of reducing or oxidizing agent present in the base.

Abstract: A method and compositions including a 1,2-dioxetane and a fluorescent compound is described. In particular, enzymatic triggering of a triggerable 1,2-dioxetane admixed with a surfactant and the fluorescent compound attached to a hydrocarbon to provide a co-surfactant in a micelle or other structure providing close association of these molecules is described. The method and compositions are useful in immunoassays and in DNA probes used for various purposes.

Abstract: Compounds having the formula: ##STR1## wherein T is a polycycloalkylidene group (e.g., adamant-2-ylidene); R is a C.sub.1-20 alkyl, aralkyl or cycloalkyl group; and Y is a fluorescent chromophore (e.g., m-phenylene), produced by reacting a compound having the formula: ##STR2## with an R-ylating agent (e.g., R.sub.2 SO.sub.4) in the presence of an alkali metal alkoxide in a polar aprotic solvent. Also, compounds having the formula: ##STR3## are produced by reacting a compound having the formula: ##STR4## with ##STR5## wherein X is an electronegative leaving group (e.g., a halogen anion such as chloride ion) in the presence of a Lewis base (e.g., a trialkyl-amine) dissolved in an aprotic organic solvent (e.g., benzene or toluene). Also, compounds having the formula ##STR6## are produced by reacting a compound of the formula ##STR7## with a tetra-O-acylated-O-hexopyranoside halide, then hydrolyzing off the protective acyl groups.