Abstract: The present invention concerns a method of enzymatically determining the pH of a specimen (e.g., a solution or a biological fluid) and a kit for conducting the method. The present method involves mixing (1) a specimen with (2) an enzyme and (3) one or more substrates for the enzyme in a buffered solution having a pH effective to provide a direct proportional relationship between the activity of the enzyme and the pH of the specimen; determining the activity of the enzyme; and correlating the activity of the enzyme to the pH of the specimen. Each of the sample, the enzyme, the substrate and the buffered solution is present in an amount effective to provide the direct proportional relationship between the activity of the enzyme and the pH of the specimen.

Abstract: The present invention concerns a method of enzymatically determining the pH of a specimen (e.g., a solution or a biological fluid) and a kit for conducting the method. The present method involves mixing (1) a specimen with (2) an enzyme and (3) one or more substrates for the enzyme in a buffered solution having a pH effective to provide a direct proportional relationship between the activity of the enzyme and the pH of the specimen; determining the activity of the enzyme; and correlating the activity of the enzyme to the pH of the specimen. Each of the sample, the enzyme, the substrate and the buffered solution is present in an amount effective to provide the direct proportional relationship between the activity of the enzyme and the pH of the specimen.

Abstract: A reflective biograting consists of an optically flat layer of a transparent composition such as silicon dioxide having a first and second surface, alternating zones of active and inactive binding reagent on the first surface, and a reflective metal layer having a thickness of at least above 1000 .ANG.. The reflective metal layer can be supported on an optically flat surface of a wafer, and the reflective metal can be aluminum, silver, gold, chromium, nickel, titanium or platinum coating on a polished wafer. Preferably, the silicon dioxide layer is formed either by direct sputtering of silicon dioxide or by coating an alkali metal silicate solution on the surface of the reflective metal, optionally containing an aminoalkylsilane and a water-soluble hydroxylated polymer such as a dextran. Alternatively, the reflective support comprises one or more reflective layer units, each reflective layer unit comprising an optically flat layer of silicon, and preferably polysilicon, on a layer of silicon dioxide.

Abstract: Novel derivatives of phencyclidine are provided as precursors for conjugating to antigenic proteins for the preparation of antibodies which bind to phencyclidine or conjugation to enzymes for use as reagents in immunoassays. The combination of the antibodies and enzyme conjugates provide a sensitive and rapid assay for phencyclidine.

Abstract: The assay of the subject invention uses DNA sequences as probes in a nucleic acid hybridization diffraction assay, to detect specific DNA sequences in a sample. Diffraction assay methodologies are applied to determine the presence and amount of analyte.This invention involves a discovery in the areas of supporting surfaces for a biogrid or biograting which provide greatly reduced non-specific hybridization and binding. A preferred process of this invention involves manufacturing a biograting for use in a light diffraction assay, and comprises adhering a uniform layer of hybridizing reagent comprising a nucleotide sequence on a smooth, solid surface and exposing the surface to UV radiation through a shadow mask with a diffraction grating pattern of lines to selectively deactivate the hybridizing reagent, leaving a biological diffraction grating design of lines of active hybridizing reagent.

Abstract: A method for identifying and quantifying chymopapain-specific IgE antibody levels in patient serum comprising binding chymopapain-specific IgE, if any, in the serum with chymopapain adhering to an insoluble support, conjugating the serum IgE with a labeled anti-IgE antibody, and measuring the level of labeled compound bound to the insoluble support or in the solution removed therefrom. Special reagents and their manufacture are also disclosed.

Abstract: A method for identifying and quantifying allergen specific IgE levels in patent serum by conjugating the IgE in the serum with allergens adhering to an insoluble support, conjugating the serum IgE with an enzyme labeled anti-IgE antibody, contacting the enzyme label with a solution of a substrate which will yield a fluorescent product in the presence of the enzyme, and measuring the level of fluorescence in the solution. Special reagents and their manufacture are also disclosed.

Abstract: A method for identifying and quantifying allergen specific IgE levels in patient serum by conjugating the serum IgE with allergens on an insoluble support, conjugating the serum IgE with an enzyme labeled anti-IgE antibody, contacting the enzyme with a solution of a substrate which will yield a fluorescent product in the presence of the enzyme, and measuring the level of fluoresence in the solution.

Abstract: In the determination of the quantity of enzyme present as a result of an immunoassay procedure, the enzyme containing component is combined with a substrate which in the presence of the enzyme yields a photometrically detectable enzymatic reaction product and a non-interfering, enzymatically inhibiting second product. The enzymatic reaction is inhibited by the addition of an inhibiting quantity of the same compound as the second product after the enzymatic reaction has continued for a time sufficient to produce photometrically detectable first products in concentrations which are a function of the quantity of enzyme present. The enzyme reaction is inhibited before the enzymatic reacton has proceeded to the point where the quantity of photometrically detectable first compound is primarily a function of the amount of substrate. This method yields a more stable test medium than methods using other enzyme inhibitors.

Abstract: A storage-stable, high potency allergenic extract is prepared by ultrafiltration, retaining fractions having molecular weights of from 1000 to 100,000, and treating the solution with gel polymer and/or carbon absorbents. The extract is dried to a moisture content of less than one weight percent water. The purified solution and aqueous reconstituted solutions of the dried extract provide a transparent, colorless solution which has greatly increased stability, remaining transparent and colorless for extended periods.

Abstract: An improved radioimmunoassay test method and device based upon competitive binding test methods wherein immunoreactions are halted at a time when the rate of change of the quantity of bound radiolabeled analyte of interest is inversely proportional to the concentration of analyte of interest in an unknown sera. Based thereon, a test device is created having a single calibration curve 36 which is accurate throughout the shelf life of the device.

Abstract: An inhibition assay for measuring the potency of allergen extracts by incubating a mixture of allergen extract and reference allergen specific IgE in a buffered solution with an insoluble support to which reference allergen is adhered. The conjugated IgE adhering to the insoluble support is reacted with an enzyme labeled anti-IgE antibody and the enzyme label is contacted with a solution of a substrate which will yield a fluorescent product in the presence of the enzyme. The level of fluoresence in the solution is measured. The percentage of inhibition of the allergen specific IgE is determined from fluorescence levels measured for various extract concentrations.

Abstract: A chymopapain derivative having the enzymatic activity reduced by at least 95% while retaining the original allergenic activity of at least 90% is suitable for skin testing to detect allergic hypersensitivity to chymopapain and for treating patients exhibiting allergic hypersensitivity to chymopapain. The enzymatic activity can be blocked by reacting thiol groups of the enzyme with and iodoacetic acid, bromoacetic acid, or a salt, ester or amide derivative thereof, under conditions which block the undesirable enzymatic activity while retaining the desired allergenic activity.

Abstract: There is provided a novel and efficent method of synthesizing 7,10-dihydro-6,11-dihydroxy-5,9,12(8H)-naphthacenetrione and analogs thereof substituted in the 1,2,3 or 4 positions. The novel method comprises reacting the appropriate quinizarinquinone with a haloprene under Diels-Alder conditions, followed by aromatization and treatment with strong acid under very mild temperature conditions. The products of this process are important intermediates in the synthesis of daunomycin and its analogs which are useful in the therapy of neoplastic ailments.

Abstract: There is provided a novel method of synthesizing certain tetracyclic quinones. In particular, there is provided a novel route to the synthesis of (.+-.)-7-deoxydaunomycinone and analogs thereof which includes the provision of novel tetrahydronaphthoquinones and tetracyclic quinone intermediates. The compounds of the present invention are provided through a route comprising a Diels-Alder addition of certain isobenzofurans to certain novel tetrahydronaphthoquinones. The products of the synthetic route provided herein may be converted into compounds of known antibiotic and antineoplastic activity.

Abstract: There is provided a novel method of synthesizing certain tetracyclic quinones. In particular, there is provided a novel route to the synthesis of (+)-7-deoxydaunomycinone and analogs thereof, which includes the provision of novel tri- and tetracyclic quinone intermediates. The -7-deoxydaunomycinone derived from naturally occurring daunomycin is a known compound, which is itself an intermediate in the preparation of the clinically accepted anti-tumor antibiotics daunomycine and its derivative adriamycin.

Abstract: There is provided a novel method of synthesizing certain tetracyclic quinones. In particular, there is provided a novel route to the synthesis of (.+-.)-7-deoxydaunomycinone and analogs thereof, which includes the provision of novel tri- and tetracyclic quinone intermediates. The -7-deoxydaunomycinone derived from naturally occurring daunomycin is a known compound, which is itself an intermediate in the preparation of the clinically accepted anti-tumor antibiotics daunomycine and its derivative adriamycin.

Abstract: There is provided a novel method of synthesizing certain tetracyclic quinones. In particular, there is provided a novel route to the synthesis of certain analogs of (+)-7-deoxydaunomycinone which includes the provision of novel tri- and tetracyclic quinone intermediates. The products of the synthetic route provided herein may be converted into compounds of known anti-neoplastic activity.