94

4778757

PATENT ABSTRACTS

M E T H O D F O R T H E D E T E R M I N A T I O N O F

S U B S T R A T E S O R E N Z Y M E A C T I V I T I E S

Shinichi Teshima, Noboru Mitsuhida, Yoshitak Nakagiri, Tsuruga, Japan assigned to Toyo Boseki Kabushiki Kaisha

There is provided an improved quantitative analysis for the determination of a substrate or enzyme. In the analysis for the enzyme, it is reac- ted with a known substrate and in the analysis for the substrate, it is reacted with a known en- zyme and then the resultant substance is reacted with an oxidase to produce hydrogen peroxide which is then reacted with a peroxidase, phenol derivatie and coupler to form a colored material which is then subjected to colorimetic analysis. The improvement resides in conducting the fore- going process by adding to the test sample a first reagent containing the oxidase, peroxidase and phenol derivative represented by the formula: See Patent for Chemical Structure (I) wherein R 1 is **a a lower alkyl group having one to five car- bon atoms or **b the group defined in **a above having a hydroxyl or sulfonic acid group, R2 is a hydrogen or halogen atom, a lower alkyl group having one to five carbon atoms, a lower acyl group having one to five carbon atoms, a lower alkylether group having one to five carbon atoms, or a lower alkoxycarbonyl group having one to five carbon atoms, said groups being unsubstituted or substituted with a hydroxyl or sulfonic acid group, and n is 0 to 4. Then, there is added to the above mixture a second reagent containing the coupler and the enzyme.

4780281

M E T H O D F O R A S S A Y O F P E R O X I D A S E E N Z Y M E O R

R E D U C I N G S U B S T R A T E A C T I V I T Y

Lawrence J Marnett, Paul E Weller assigned to Wayne State University

A preferred method wherein 5-phenyl-4- pentenyl-hydroperoxide (PPHP) is reduced to 5- phenyl-4-pentenyl-alcohol (PPA) by plant and animal peroxidases in the presence of reducing substrates is described. The method also uses related homologs containing 3 to 8 carbon atoms. PHP and PPA are rapidly isolated with solid phase extraction, separated by isocrated

reverse phase high performance liquid chromatography, and quantitated with a fixed- wavelength ultraviolet detector. The procedure described in suitable for detecting peroxide reducing enzymes, determining the kinetic pro- perties of heme- and non-heme-containing peroxidases, and evaluating oxidizable com- pounds as reducing substrates for peroxidases. The method identifies compounds which are reducing substrates and also ranks them for rela- tive activity. The method can be used to identify active antithrombotic, antimetastatic, or anti- inflammatory drugs as substrates as welt as detect and characterize mammalian peroxidases.

4780405

C A R B O N I C A N H Y D R A S E I N H I B I T O R - T A G G E D N U C L E I C

A C I D P R O B E S

Emil T Kaiser, Gary F Musso, Soumitra Ghosh, Leslie E Orgel, Geoffrey Wahl assigned to Siska Diagnostics Inc

Nucleic acid hybridization probes are provided which comprise nucleoside bases or terminal nucleotide phosphates chemically linked to aromatic sulfonamide inhibitors of carbonic an- hydrase. Methods of preparing probes of the in- vention, intermediates used in such methods, and methods of using the probes of the invention in hybridization assays are also provided. A probe of the invention is detected by binding to it a reporter group, such as a homopolymer or heteropolymer of enzymes, which includes a car- bonic anhydrase which binds to the inhibitor linked to the probe, and then detecting the bound reporter group, as by production of a fluorescent or colored product in a reaction cata- lyzed by an enzyme component of the reporter group. Also provided are enzyme immunoassays wherein detection of antibody is by a process which comprises a chemical reaction catalyzed by a carbonic anhydrase.

4780406

F L O W C Y T O M E T R I C M E A S U R E M E N T O F T O T A L D N A

A N D I N C O R P O R A T E D H A L O D E O X Y U R I D I N E

Frank A Dolbeare, Joe W Gray assigned to The Regents of the University of California

A method for the simultaneous flow cytometric