Molecular distribution of elastase between its two main inhibitors: direct quantitation of elastase-alpha 2-macroglobulin complex with a novel ELISA technique.
A novel enzyme-linked immunosorbent assay for the quantitation of elastase linked to alpha 2-macroglobulin (alpha 2M) (elastase-alpha 2M complex, EMC) in body fluids is presented. The assay has a lower detection limit of 1.5 ng bound elastase per ml. The critical factor allowing immunological detection of alpha 2M-bound elastase is the addition of phenyl methyl sulphonyl fluoride (PMSF) to the assay buffer. The assay has been used to quantitate EMC in plasma and serum of healthy volunteers, and assess the distribution of elastase between its two main inhibitors, alpha 2M and alpha 1-proteinase inhibitor (API). EMC levels in serum samples from volunteers were significantly higher than in plasma (26.9 vs 21.9 ng/ml, p = 0.05, Student's t-test for paired samples). The API-bound elastase levels were 70.2 and 170.1 ng/ml in plasma and serum respectively (statistically significantly higher in serum, p less than 0.0001). The ratio of macroglobulin to alpha 1-proteinase-bound elastase levels was 0.333 in plasma and 0.168 in serum (p less than 0.0001). As alpha 2M bound elastase retains some proteolytic activity, direct measurement of circulating levels in inflammatory conditions should be of interest.
Department of Medicine, Royal Postgraduate Medical School, London, UK.
E O
EO
Adeyemi
H J
HJ
Hodgson
D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000431:Ethanol; D006801:Humans; D010196:Pancreatic Elastase / Q000097:blood*; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D010949:Plasma / Q000032:analysis; D000515:alpha 1-Antitrypsin / Q000378:metabolism*; D000511:alpha-Macroglobulins / Q000378:metabolism*
D000515:alpha 1-Antitrypsin; D000511:alpha-Macroglobulins; D000431:Ethanol; D010664:Phenylmethylsulfonyl Fluoride; D010196:Pancreatic Elastase