Enzyme immunoassay of human placental and germ-cell alkaline phosphatase in serum

Abstract

Placental alkaline phosphatase (placental ALP, PLAP) and germ-cell ALP (GCAP, also known as placental-like ALP), expressed in gonadal cancer tissues, are potential tumor markers. Four monoclonal antibodies, raised against PLAP and recognizing different epitopes, were selected to study the influence of the following variables on the accuracy of PLAP and GCAP measurement: phenotype, molecular form, and glycation pattern of PLAP and GCAP; incubation temperature; and interferences by serum during immunobinding. Nine GCAP phenotypes were identified, interacting with each antibody at a lower affinity than was seen for the more common PLAP phenotypes. Antibody affinity is higher for the free hydrophilic dimeric forms of PLAP and GCAP, and is not influenced by the degree of glycation. In serum or tissue extracts, measurement of PLAP or GCAP is most nearly accurate when immunoincubations are performed at 37 °C, with use of antibodies 327 and 7E8, respectively. In addition, correct measurements are achieved only when, during immunobinding, serum is incubated with an equal volume of deoxycholate (9 g/L final concentration).