Rapid quantitative, specific measurement of pancreatic amylase in serum with use of a monoclonal antibody.
In this rapid quantitative assay for pancreatic alpha-amylase (EC 3.2.1.1) in serum, we precipitate salivary amylases by 10-min incubation with monoclonal anti-salivary amylase antibody immobilized on particles of polyvinylidene fluoride. We then centrifuge the serum mixture and measure the pancreatic amylase activity remaining in the supernate by a kinetic method. The assay requires 50 microL of serum and the standard curve is linear to at least 1300 U of pancreatic amylase per liter of serum. CVs were 1.3% within-run, 6-8% day-to-day. Apparent analytical recovery of pancreatic amylase activity added to serum was 101% +/- 2%. Addition of purified salivary amylase, 356 U/L, to sera gave a value for apparent pancreatic amylase of less than 4 U/L, or 1% of the added salivary amylase activity. This assay correlated well with an electrophoretic method (slope, 0.97-0.99; intercept, 0.5 to -4 U/L; correlation coefficient, 0.946-0.990; and standard error of the estimate 3-5 U/L). Estimated normal reference intervals with maltotetraose as substrate were: total amylase, 39-118 U/L; pancreatic amylase, 11-50 U/L; and salivary amylase, 18-79 U/L.
D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007700:Kinetics; D008320:Maltose / Q000031:analogs & derivatives / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D012015:Reference Standards; D012463:Saliva / Q000201:enzymology; D000516:alpha-Amylases / Q000097:blood*
D000911:Antibodies, Monoclonal; D009844:Oligosaccharides; D008320:Maltose; C009819:maltotetraose; D000516:alpha-Amylases