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High throughput Diethylaminoethyl cellulose chromatographic purification of Alkaline Phosphatase from Hepatopancreas of Brown shrimp, Metapenaeus monoceros.

Alkaline phosphatase was isolated from Brown shrimp hepatopancreas by homogenisation at 3,000 rpm for 10 min at 4 o C, clarification at RCF of 1681.1 x g for 5 min at 4 o C, and precipitation at 65% ammonium sulfate saturation level at 0 o C. Alkaline phosphatase purification was optimised in DEAE-cellulose columns using binding buffer of pH 7.6, 8.0, 8.4, or 8.8 and ionic strength of 0.00, 0.05, 0.10, or 0.15 M NaCl, and elution buffers of gradient 0.10-0.25, 0.10-0.35, 0.10-0.45 or 0.10-0.55 M NaCl in 25 min and flow rate of 0.5, 1.0, 1.5, or 2.0 mL/min. Binding buffers of pH 8.4 and ionic strength 0.1 M NaCl at flow rate of 1 mL/min is suitable for optimal binding of alkaline phosphatase to the column, and elution buffer of ionic gradient of 0.10-0.35 M NaCl in 25 min at a flow rate of 1.5 mL/min is suitable for optimal resolution.