Solid phase immunoassay for high molecular weight alkaline phosphatase in human sera using a specific monoclonal...

A murine hybridoma producing monoclonal antibody (MoAb HMAP-1) to human high molecular weight alkaline phosphatase (HMAP) was derived using enzyme partially purified from human serum as the immunogen. The antibody did not cross-react with the liver, bone, intestinal or placental isozymes of alkaline phosphatase (AP), and did not react with AP in bile. A monoclonal antibody immunocatalytic assay for HMAP in serum was developed using MoAb HMAP-1 bound to nitrocellulose membrane discs. The method was rapid and reproducible, giving good correlation with results from cellulose acetate membrane electrophoresis and having the added advantages of increased sensitivity and specificity. A large number of sera can now be assayed to evaluate the significance of serum HMAP in relation to other AP activities and to the differential diagnosis of liver disease.