Coulter Corporation

Coulter Electronics, Inc.

Coulter International Corp.

Gerald E Jaffe

Frank J Lucas

James H Carter

Steven E Bott

Marilyn M Solorzano

Gary A Mitchell

Harold R Crews

Mathew M Madappally

Giovanni Bucolo

Nazle Kuylen

Francis J Lucas

Nancy M Garcia

Maria Barcelon

The necessity to detect blast cells in a patient sample to determine a treatment regime for leukemia has long been desired. The present invention relates to a method and assay reagents for identification of blast cells in a fluid sample. The method further enables the determination of the lineage of the blast cells. The new method provides a determination of blast cells in less time than prior art methods.

Identification of blast cells in a leukocyte cell preparation

An assay compound or a salt thereof for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound as well as methods of using these compounds to measure enzyme activity are also disclosed.

Method for determining activity of enzymes in metabolically active whole

Assay reagent

Method of making an assay compound

A method for assaying the activity of an enzyme inside a metabolically active whole cell is disclosed. The assay compound includes a leaving group and an indicator group. The leaving group is selected from the group comprising amino acids, peptides, saccharides, sulfates, phosphates, esters, phosphate esters, nucleotides, polynucleotides, nucleic acids, pyrimidines, purines, nucleosides, lipids and mixtures thereof. The indicator group is selected from compounds which have a first state when joined to the leaving group, and a second state when the leaving group is cleaved from the indicator group by the enzyme. Preferably, the indicator compounds are rhodamine 110, rhodol, and fluorescein and analogs of these compounds. A method of synthesizing the compound is also disclosed.

Water soluble xanthylium derivative substrates of rhodamine 110 and rhodol permit spectrophotometric and fluorescent measurements of trypsin-like enzymes without the addition of organic solvent additives and/or special water solubilizing agents. These novel substrates exhibit increased sensitivity for determining low levels of activity of trypsin-like enzymes such as proteolytic enzymes, cofactors, activators, antiactivators, and inhibitors. These substrates can be substituted for fibrinogen to monitor the pathways of blood coagulation.

Water soluble zanthylium derivative substrates

Water soluble xanthylium derivative substrates of rhodamine 110 and rhodol permit spectrophotometric and fluorescent measurements of trypsin-like enzymes without the addition of organic solvent additives and/or special water solubilizing agents. These novel substrates exhibit increased sensitivity for determining low levels of activity of trypsin-like enzymes such as proteolytic enzymes, cofactors, activators, antiactivators, and inhibitors. These substrates can be substituted for fibrinogen or monitor the pathways of blood coagulation.

Water soluble xanthylium derivatives substrates

A colorimetric method for the determination of gamma-glutamyl transpeptidase enzyme activity in a biological fluid comprises catalytically reacting the enzyme in the fluid with a substrate of a water-soluble salt of a compound selected from the group consisting of L-gamma-glutamyl-3-sulfoanilide and L-gamma-glutamyl-4-sulfoanilide in aqueous solution to produce the corresponding aniline sulfonic acid salt as a cleavage product, diazotizing the cleavage product, and coupling the diazotized product with a coupling compound to produce a dye having a color intensity proportional to the enzyme activity.

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Pembroke Pines, FL, United States of America

Gerald E Jaffe