New immunocapture enzyme (ICE) assay for quantification of cancer procoagulant activity: studies of inhibitors.
A new, sensitive and specific immunocapture enzyme (ICE) assay for quantitation of the enzymatic activity of cancer procoagulant (CP) has been developed. The assay had good reproducibility (inter- and intra-assay CV were 6.4% and 5.7% respectively) and was linear for concentrations of CP from 0.5 microgram/ml to 10 micrograms/ml (r2 = 0.995). Using this assay the inhibition of CP by iodoacetamide, mercuric chloride, E-64, leupeptin and antipain was demonstrated. There was no significant effect of cystatin and natural plasma proteinase inhibitors alpha 1-antitrypsin, alpha 1-antichymotrypsin, alpha 2-macroglobulin and antithrombin-III/heparin, on the activity of the CP.
Mielicki
W P
WP
University of Colorado Health Sciences Center, Denver 80262.
Tagawa
M
M
Gordon
S G
SG
D014408:Biomarkers, Tumor / Q000097:blood; D002863:Chromogenic Compounds; D003546:Cysteine Endopeptidases / Q000097:blood* / Q000187:drug effects / Q000276:immunology; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D009363:Neoplasm Proteins; D009369:Neoplasms / Q000097:blood; D012680:Sensitivity and Specificity
D014408:Biomarkers, Tumor; D002863:Chromogenic Compounds; D015853:Cysteine Proteinase Inhibitors; D009363:Neoplasm Proteins; D003546:Cysteine Endopeptidases; C030910:cancer procoagulant