Solid-phase radioimmunoassay for human neutrophil elastase: a sensitive method for determining secreted and cell-associated enzyme.
A solid-phase radioimmunoassay for measuring neutrophil elastase in the range 0.08-4 ng/ml has been developed. A monospecific, precipitating antibody capable of inhibiting elastinolysis was produced by repeated immunizations of a goat. The IgG fraction and affinity-purified antibodies of this serum were then obtained and used to develop this radioimmunoassay. There was no cross-reactivity in binding of the radiolabeled antisera with lactoferrin, cathepsin G, or serine proteinases with amino-terminal amino acid sequence homology. Although serum influences the measurement of catalytically active neutrophil elastase when compared to diisopropylfluorophosphate-treated neutrophil elastase, antigenic elastase may still be measured in body fluids. Furthermore, this assay is more sensitive than commercially available substrates used for quantitating neutrophil elastase by functional activity. We have found this quantitative assay extremely useful in balance studies to measure secreted and cell-associated elastase and in screening of biological fluids for the presence of the enzyme.
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D004705:Endocytosis; D006041:Goats; D006801:Humans; D007457:Iodine Radioisotopes; D008297:Male; D008832:Microchemistry; D009504:Neutrophils / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000097:blood* / Q000378:metabolism; D011863:Radioimmunoassay / Q000379:methods; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D015054:Zymosan / Q000494:pharmacology
D007457:Iodine Radioisotopes; D012710:Serum Albumin, Bovine; D015054:Zymosan; D010196:Pancreatic Elastase