Affinity elution from a phosphonic acid-Sepharose derivative in the purification of human liver alkaline phosphatase.
The compound p-aminobenzylphosphonic acid has been coupled via an azo linkage to tyraminyl-Sepharose 4B. This derivative at pH 6.0 bound most of the protein and all of the alkaline phosphatase in a crude preparation from human liver. The phosphatase was selectively eluted with the substrate 2-naphthylphosphate and a purification of 400-fold obtained. This step, when incorporated into a procedure for the purification of human liver alkaline phosphatase, yielded essentially pure enzyme.
D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D006801:Humans; D008099:Liver / Q000201:enzymology*; D063065:Organophosphonates / Q000138:chemical synthesis; D011134:Polysaccharides / Q000031:analogs & derivatives / Q000138:chemical synthesis; D012685:Sepharose / Q000031:analogs & derivatives / Q000138:chemical synthesis
D063065:Organophosphonates; D011134:Polysaccharides; D012685:Sepharose; D000469:Alkaline Phosphatase