Detection of Candida proteinase by enzyme immunoassay and interaction of the enzyme with alpha-2-macroglobulin.
Double antibody immunoassay of secretory aspartic proteinases from 2 strains of the yeast Candida albicans was performed in microtest plates with rabbit antibody and peroxidase conjugated rabbit antibody. The test detects approximately 0.1 ng of proteinase and is 2 orders of magnitude more sensitive than direct measurement of enzymic activity with hemoglobin as a substrate. The immunoassay was affected both by alkaline denaturation, which is a common feature of aspartic proteinases, and by the presence of the proteinase scavenger alpha-2-macroglobulin. Conditions are described that allow for comparison of proteinases from different strains of yeast and minimize the interference of macroglobulin.
D000818:Animals; D002176:Candida albicans / Q000201:enzymology*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D007122:Immunoelectrophoresis; D007124:Immunoenzyme Techniques; D010447:Peptide Hydrolases / Q000032:analysis* / Q000276:immunology; D000511:alpha-Macroglobulins
D007106:Immune Sera; D000511:alpha-Macroglobulins; D010447:Peptide Hydrolases