Gds Technology, Inc.

Aurora F deCastro

Surendra K Gupta

Steven M Shantz

Arun K Agarwal

The present invention provides a kinetic or rate method for the determination of cholesterol in free or bound form with of a cholesterol oxidase enzyme obtained from Bacterial strain NRRL B-18713 which inherently has a high Michaelis and Menton, Km, of about 10-2 to 10-3 M for cholesterol and optionally, of cholesterol esterase. The method can measure oxygen consumption, hydrogen peroxide or cholestenone formed. The present invention also provides a test composition for the kinetic determination of cholesterol, comprising cholesterol oxidase with an inherently high Km of about 10-2 to 10-3 M for cholesterol and optionally cholesterol esterase and a system for the determination of hydrogen peroxide, cholestenone or oxygen consumption.

Method, composition and device for the determination of cholesterol

The fluid transfer device of the present invention is used for example, to provide access to a vacuum tube which is sealed with a stopper. The device has a cylindrical body with a top wall portion and first and second conduits fixed and extending therethrough. The first, or pouring conduit, and second, or pressure equilibrating conduit, extend generally in a parallel manner above and below the top wall portion, and their respective bottom portions are contained within the body of the hollow device. The pair of conduits are constructed and arranged within the body of the device to pierce and enter through the stopper of the sealed container to permit the delivery of fluid. Further, the device has a structure that can be easily and economically manufactured for disposable use.

Fluid transfer device

The present invention provides a new methodology and test composition for determining the presence of theophylline in test samples. The methodology employs enzymes that utilize or recognize theophylline as a substrate to measure the concentration thereof in samples, including body fluids. This new approach utilizes enzymes as opposed to traditional methods which use antibodies for the recognition of theophylline. The enzymatic approach to theophylline determination is quick, simple and convenient and allows test systems to be made in liquid as well as in dry-chemistry formats. Various protocols, systems or methodologies may be used for assaying and relating the results to the amount of theophylline present. Methods for obtaining theophylline utilizing or recognizing enzymes are also described.

Enzymatic determination of theophylline

Arylacylamidase can be stabilized by inhibiting conformational changes using either o-cresol or benzoic acid or salts of benzoic acid as a stabilizing agent. The compositions show significantly enhanced stability of arylacylamidase in aqueous solution, lyophilized, and solid-phase formats.

Method of preserving arylacylamidase in aqueous solution

A method and unitized test composition is described for the estimation of an anilide in which the enzymatic hydrolysis of the anilide and colorimetric quantitation of aniline or aniline derivative can be done simultaneously. The hydrolysis of the anilide is catalyzed by a known enzyme, arylacylamidase, E. C. 3.5.1.13. Stabilization of the enzyme is provided by the addition of controlled amounts of a compound containing alcoholic and/or aromatic groups such as ortho-cresol, isopropanol or benzoate. Basically, the unitized test composition comprises (i) arylacylamidase, (ii) a controlled amount of an organic compound containing alcoholic and/or aromatic groups which acts as both a stabilizer for the arylacylamidase and forms a colored product with aniline, and (iii) a novel oxidant/catalytic agent for accelerating color development. The benefit of such methodology results in a one step addition of the complete reagent to the sample, serum or matrix containing the anilide as opposed to a several step addition. The consequence of this is that only one reagent channel is required to perform the test in an automated analyzer as opposed to the usual requirement that several channels be utilized. This method also makes possible the development of a solid-phase test device such as a dip and read test strip containing all the reagents necessary for the test of an anilide. In addition, a method for the stabilization of arylacylamidase enzyme is described.

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Union, MI, United States of America

Aurora F deCastro