title: Purification of human adult and foetal intestinal alkaline phosphatases by monoclonal antibody immunoaffinity chromatography.
abstract: We have used the technique of monoclonal antibody immunoaffinity chromatography to purify adult and foetal intestinal alkaline phosphatases. Pure adult intestinal enzyme was obtained from a crude tissue extract with a single immunoaffinity chromatographic step in yields exceeding 95%. An additional ion-exchange chromatographic step was necessary for purification of the foetal enzyme, but yields still exceeded 70%. Experiments to optimize the efficiency of the monoclonal antibody immunoaffinity chromatography procedure suggest that the relative strength of binding of an antibody to its antigen is the most important factor to consider when constructing such columns. A column made from an antibody of too low an avidity will not retain the enzyme, while one of too high an avidity will make elution of enzyme in the active state difficult. A scheme is suggested for the application of this technique to a general approach to enzyme purification.
mesh_terms: D000328:Adult; D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007163:Immunosorbent Techniques; D007422:Intestines / Q000196:embryology / Q000201:enzymology*; D007527:Isoenzymes / Q000302:isolation & purification*; D008970:Molecular Weight
chemical_list: D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D000469:Alkaline Phosphatase