Kimberly A Hines

US Patent:

20020192736, Dec 19, 2002

Filed:

Feb 23, 2001

Appl. No.:

09/792741

Inventors:

Paul Davis - Dublin OH, US
Kimberly Hines - Crystal Lake IL, US

Assignee:

Pierce Chemical Company

International Classification:

C12Q001/26
C09K003/00

US Classification:

435/025000, 252/183120

Abstract:

A composition for assaying of peroxidase activity: (a) at least one chemiluminescent cyclic diacylhydrazide, (b) at least one azine enhancer, and (c) at least one oxidizing agent wherein said azine enhancer is present, with respect to the total molar amount of azine compounds in the composition, as less than 0.005 moles of azine compounds of said total molar amount of azine compounds of azine enhancer having a hydrogen atom bonded to a nitrogen atom of the azine ring. The presence of larger amounts of azine enhancer having hydrogen atom bonded to the nitrogen atom of the azine ring has been found to poison or deteriorate the quality and efficiency of the chemiluminescent effect.

US Patent:

6432662, Aug 13, 2002

Filed:

May 20, 1999

Appl. No.:

09/171334

Inventors:

Paul D. Davis - Dublin OH
Kimberly Hines - Crystal Lake IL

Assignee:

Pierce Chemical Company - Rockford IL

International Classification:

C12Q 128

US Classification:

435 28, 435 8, 435 27

Abstract:

Chemiluminescent detection of molecules of synthetic or natural origin such as proteins and nucleic acids (DNA and RNA), as well as other biologic molecules, is increasingly replacing radioactive detection as the method of choice where sensitivity is critical. In such assays, luminescence is customarily achieved by the oxidation of a luminol or isoluminol substrate in the presence of an oxidizing agent such as hydrogen peroxide or hydrogen peroxide source, such as perborate, and a peroxidase catalyst such as horseradish peroxidase. To obtain useful levels of luminescence (e. g. , detectable levels) by customary techniques, a luminescent enhancer is also present during oxidation. It has been found in the practice of the present invention that azine enhancers have contained an impurity which reduces the properties of the chemiluminescent assay working solutions. The present invention describes a working solution, a process of using the working solution, and kits containing solutions which can be mixed to form the working solutions comprising a solution which is useful for the chemiluminescent assay of peroxidase activity comprising: a) at least one chemiluminescent cyclic diacylhydrazide, b) at least one azine enhancer, and c) at least one oxidizine agent wherein said azine enhancer comprises less than 0. 005 parts mole/mole total basis of azine compounds having a hydrogen atom bonded to a nitrogen atom of the azine ring.