Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving oxidoreductase
Patent
1991-11-08
1999-02-09
Leary, Louise N.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving oxidoreductase
435 71, 435 4, 435 23, 435 18, 435 794, 435 793, 435 792, 530356, 530354, C12Q 128, C12Q 137, G01N 3353
Patent
active
058692770
ABSTRACT:
The present invention relates to the development of a modified substrate capture immunoassay useful in the detection of a neutral metalloproteinase enzyme, type IV collagenase. Capture and immobilization of this enzyme can be achieved by coating the microtiter plate with gelatin, an alternative substrate for this enzyme. The captured enzyme can then be detected by affinity purified rabbit anti-type IV collagenase antibodies prepared against synthetic peptides from the amino terminus of the enzyme. Soluble type IV collagenase can readily be detected in samples known to contain this enzyme. Using purified enzyme this assay method can detect less than 50 ng of latent type IV collagenase. Using EDTA, an inhibitor of this metalloproteinase, gelatin binding can be shown to be independent of catalytic activity.
REFERENCES:
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patent: 4677058 (1987-06-01), Tryggvason et al.
patent: 4804625 (1989-02-01), Morrison et al.
Lydia Woods Schindler, "Understanding the Immune System," NIH Pub. No. 92-529, p. 36 (U.S. Dep't of Health & Human Servs. Oct. 1991).
Robertson et al; A sensitive Microplate Assay for Detection of Proteolytic Enzymes Using Radiolabeled Gelatin, Analytical Biochem 172, pp. 284-287 (1988). Month not available.
Liotta Lance
Stetler-Stevenson William G.
Leary Louise N.
Rucker Susan S.
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