Appeal No. 96-1355
Application 07/896,209
bringing the peroxide and indicator in the assay medium into contact
with the released myeloperoxidase in that area of the membrane where the
myeloperoxidase is released . . .
Free is directed to methods and indicator compositions for “detecting erythrocytes
by their hemin content when present in a body fluid also containing leukocytes with their
peroxidase content” and for “detecting leukocytes by their peroxidase content when
present in a body fluid also containing erythrocytes with their hemin content.” The
reference teaches that “various catalytically active substances (substances having
peroxidative activity) can be differentiated under a given set of conditions.” That is, the
peroxidase activity of the leukocytes can be distinguished from the peroxidative activity of
hemin in the erythrocytes, and vice versa, without separation, by “careful adjustment of
peroxide concentration, pH and indicator concentration and choice of indicator.” See
column 1, lines 52-69, and column 3, lines 1-10.
Meiattini discloses “a reagent suitable both for the detection and for the quantitative
determination of leukocytes in biological samples through the measurement of
[myeloperoxidase] activity . . . having such a specificity that hemoglobin does not bring
about interferences at all even in the presence of several erythrocytes . . .” See page 3.
Fetter separates whole blood into red cell and colorless fractions so that
chromogenic indicators can be used to detect analytes in the colorless fraction without
interference from the intensely colored red cells.
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