Time-resolved immunofluorometric assay of human pancreatic isoamylase in serum, with use of two monoclonal antibodies.
This new method for determining pancreatic isoamylase (EC 3.2.1.1) in serum involves two monoclonal antibodies: one immobilized in a microtitration well (the capture antibody), the other biotinylated. After the sample is incubated with the two antibodies, the captured immunocomplex is quantified by adding streptavidin labeled with a europium chelator and measuring the specific Eu3+ fluorescence in a time-resolved mode. Three assay protocols are proposed, involving incubation times of 90, 45, or 25 min. The assay has low (0.005%) cross-reactivity with the salivary isoenzyme. Analytical performance was satisfactory. Results correlate well with results obtained by measuring total amylase activity or by measuring pancreatic isoamylase activity after immunoinhibition. Unlike numerous current amylase assays, this method measures enzyme mass rather than enzyme activity. Potentially, this is a highly specific assay.
Diamandis
E P
EP
CyberFluor Inc., Toronto, Ontario, Canada.
Papanastasiou-Diamandi
A
A
Lustig
V
V
Khosravi
M J
MJ
Tan
A
A
D000328:Adult; D000681:Amylases / Q000097:blood; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005063:Europium; D005455:Fluorescent Antibody Technique; D006026:Glycoside Hydrolases / Q000097:blood*; D006801:Humans; D007201:Indicator Dilution Techniques; D007517:Isoamylase / Q000097:blood*; D008875:Middle Aged; D010179:Pancreas / Q000201:enzymology*; D012016:Reference Values; D019809:Streptavidin
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005063:Europium; D019809:Streptavidin; D000681:Amylases; D006026:Glycoside Hydrolases; D007517:Isoamylase