P-178: Separation and Identification of Alkaline Phosphatase Isozymes during Pregnancy

Background: Alkaline phosphatase (ALP), (EC 3.1.3.1) is a hydrolase enzyme responsible for removing phosphate groups from various molecules in the body. In humans, ALPis present in all tissues such as liver, bile duct, kidney, bone, and the placenta which detection of its activity is so useful in molecular biology. Pregnancy is associated with normal physiological changes that assist fetal survival as well as preparation for labor. The aim of this research was evaluating the role of various ALP isozymes in the first trimester abortion. Materials and Methods: Enzyme Assay: Blood samples were collected from (60) pregnant and (30) aborted women who admitted in Rasht hospital. ALPase activity was measured by using Stopped Spectrophotometric method. The change in absorbance at 410 nm was determined that is indicative of enzyme activity. Total Protein Determination: Protein concentration was measured by bradford method and absorbance determined at 595nm using bovine serum albumin (BSA) as standard. Separation of ALP lsoenzymes by Gel electrophoresis and Partial Purification of Human ALPase: Using SDS-PAGE, various isoenzymes of ALP were detected in serum of our participant. The collected samples were subjected to different steps of purification including precipitation by ammonium sulphate (NH4)2SO4, dialysis and ion-exchange chromatography. Results: It was found that ALP in serum of abort person had fewer isozymes in comparison with the pregnant group (p<0.05). Conclusion: Although assay of ALP activity in serum is a common clinical test, the physiological function of this enzyme remains uncertain. ALP activity: This article will focus on biochemical results to pregnancy, which fall into 2 different categories. On the basis of our results we conclude that isozymes of ALP in serum are a diagnostic marker for pregnant women in high risk for early abortion.