Purification of human serum paraoxonase: A simple and rapid method

Abstract


A. J. Mahadesh Prasad*, K. Kemparaju, Elizabeth A. Frank and Cletus J. M. Dâ??Souza

Paraoxonase/arylesterases (EC.3.1.8.2) is an enzyme found tightly associated with high density lipoprotein particle in serum. Because of its unique enzyme activity, antioxidant property and its role as an anti atherosclerotic molecule, various methods are used for its purification from human serum. Methods involved in its purification are elaborate and complicated. Also the yield and final activity are highly variable. Here, we report a 2 step method of purification involving affinity chromatography on cibacron blue sepharose followed by gel filtration on sephadex G50.The final preparation was 27.7 fold purified compared with the serum and gave a single band in SDS-PAGE by silver staining.

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