The Role and Efficiency of Ammonium Sulphate Precipitation in Purification Process of Papain Crude Extract

Abstract

It has been common to do fractionation (for example using ammonium sulphate as a precipitating agent) before doing a more sophisticated method for purification of a protein. The logic behind this is easy to understand, but in fact, the precipitation step often causes severe loss in yield and activity of the protein, making the whole purification effort too costly. In this work we evaluated the specific activity (thus, purification factor) and total activity (yield) during the purification process of papain from a crude extract using ion exchange chromatography (IEC), with and without prior fractionation using ammonium sulphate. Detail assays in each step were recorded and SDS-PAGE was also done to reveal the protein profile of the purification products.