Development of a reference material for alkaline phosphatase.
In developing a Reference Material for alkaline phosphatase, we studied the stability, kinetic properties, and commutability of separate preparations of the purified enzyme from human liver, intestine, bone, and placenta. The Michaelis constants (Km) for the preparations from liver, bone, and intestine agreed well with the Km values we obtained for five human serum specimens, whereas that for the placental isoenzyme differed significantly. The first three isoenzymes exhibited nearly identical response-surface patterns, which closely paralleled those observed for 12 human serum specimens (commutability), but not that of the placental isoenzyme. Thus, we believe that a reference material could equally well consist of either the bone, intestinal, or liver isoenzyme. All four isoenzymes were satisfactorily stable in temperature-accelerated degradation studies. We chose the liver isoenzyme as an appropriate reference material because liver tissue is easier to obtain than bone or intestine and the isoenzyme is abundant in liver, is easy to extract, and is the one most commonly increased in human serum. This material is stable at -20 degrees C, is free of interfering and degradative enzymes and, being of human origin, is commutable with the enzyme in human serum.
D000328:Adult; D000469:Alkaline Phosphatase / Q000302:isolation & purification / Q000592:standards*; D000704:Analysis of Variance; D001842:Bone and Bones / Q000201:enzymology; D002648:Child; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007422:Intestines / Q000201:enzymology; D007527:Isoenzymes / Q000302:isolation & purification / Q000592:standards*; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D010920:Placenta / Q000201:enzymology
D007527:Isoenzymes; D000469:Alkaline Phosphatase