Minnesota Mining and Manufacturing Company

Adeza Biomedical Corporation

Biowhittaker, Inc.

Axionics, Inc.

Axonica, Inc.

Emanuel Calenoff

Yuh-Geng Tsay

Ruth M Jones

Myron A Beigler

John R Scott

James L Nichols

Gerald L Friesen

Rick P Trebino

Eric K Gustafson

John Lee

Scott, John R

Tsay Yuh-Geng

Ruth M Johnson

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. The smooth, solid surface is preferably selected from the group consisting of polysilicon and single crystalline silicon surfaces. The diffraction hybridizing assay method of this invention for determining the presence or quantity of an analyte in an aqueous sample comprises contacting a nucleic acid sequence diffraction biogrid with the sample under proper circumstances and for a sufficient time to permit nucleic acid hybridization between a nucleic acid sequence probe and an analyte; separating the biogrid from the sample; illuminating the biogrid with light from a light source; and determining the light diffracted by the diffraction hybridization assay surface.

DNA probe diffraction assay and reagents

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.

Fluorometric assay of chymopapain hypersensitivity and reagents therefor

A storage-stable, high potency allergenic extract is prepared by ultrafiltration, retaining fractions having molecular weights of from 1000 to 100,000, and drying the retained fraction to a moisture content of less than one weight percent. The extract can also be pretreated with amylase before ultrafiltration, treated with affinity chromatography before drying, and/or treated with gamma radiation after drying.

Stable antigenic extracts methods

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.

Assaying allergen specific IgE levels with fluorogenic enzyme labeled

A method for identifying and quantifying total IgE levels in patient serum by conjugating the IgE in the serum with anti-IgE antibody 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 fluoresence in the solution. Special reagents and their manufacture are also disclosed.

Assaying total IgE levels with fluorogenic enzyme labeled antibody

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.

Fluorometric assay of allergic reactions

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.

Stable allergenic extracts and methods

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.

Fluorometirc enzyme inhibition immunoassay for measuring potency of

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.

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Burlingame, CA, United States of America

Emanuel Calenoff