Separation of alkaline phosphatase isoforms with and without intact glycan-phosphatidylinositol anchors in aqueous polymer phase systems.
Alkaline phosphatase (ALP) isoforms can be distinguished from each other by their partition characteristics in aqueous two-phase systems composed of water-soluble polymers, the phases of which are differentially sensitive to the presence of glycan-phosphatidylinositol anchors. Compared with detergent-based systems, the aqueous polymer systems have the advantage that micelle formation does not take place. Partition of anchor-intact and anchor-degraded molecules in the latter systems is further improved by attachment of a hydrophobic ligand to one of the phase forming polymers. In this way, anchor-intact molecules can be separated from molecules with degraded anchors in a single partition step. The method has been used to confirm that ALP in human serum is predominantly anchor degraded, whereas in bile it retains its anchor intact.
Raymond
F D
FD
Department of Chemical Pathology, Royal Postgraduate Medical School, London, UK.
Moss
D W
DW
Fisher
D
D
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods*; D003911:Dextrans / Q000737:chemistry*; D017261:Glycosylphosphatidylinositols / Q000737:chemistry*; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification; D011092:Polyethylene Glycols / Q000737:chemistry*; D014867:Water
D003911:Dextrans; D017261:Glycosylphosphatidylinositols; D007527:Isoenzymes; D014867:Water; D011092:Polyethylene Glycols; D000469:Alkaline Phosphatase