Design and applications of biomimetic anthraquinone dyes. Purification of calf intestinal alkaline phosphatase with immobilised terminal ring analogues of C.I. reactive blue 2.
A 330-fold one-step purification of alkaline phosphatase from a crude calf intestinal extract has been achieved using specific elution with inorganic phosphate (5 mM) from a purpose designed adsorbent comprising a terminal ring phosphonate analogue of C.I. Reactive Blue 2 coupled to Sepharose CL-6B-200. The resulting alkaline phosphatase preparation displayed a specific activity in excess of 1000 U/mg and was of equivalent purity to commercial "high purity" preparations as deduced by sodium dodecyl sulphate polyacrylamide gel electrophoresis and specific activity comparisons.
Lindner
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Unilever Research, Colworth Laboratory, Sharnbrook, Bedford, U.K.
Jeffcoat
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Lowe
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D000469:Alkaline Phosphatase / Q000302:isolation & purification*; D000818:Animals; D000880:Anthraquinones; D002417:Cattle; D002853:Chromatography, Liquid; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D007422:Intestines / Q000201:enzymology*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D014227:Triazines
D000880:Anthraquinones; D004396:Coloring Agents; D007202:Indicators and Reagents; D014227:Triazines; D012710:Serum Albumin, Bovine; C012842:Cibacron Blue F 3GA; D000469:Alkaline Phosphatase