Partition coefficient studies in integrated aqueous two phase (ATPi) extraction with free tri-azine dye ligands for papain partitioning from its crude latex

Integrated aqueous two phase extraction (ATPi) system- an emerging technique for purification of proteins in single step is widely used in downstream processing, for this method yields proteins with high purity and is cost effective. The study is based on affinity dye based aqueous two phase extraction system for papain partitioning from its crude latex. Response surface methodology using central composite design approach is one of the promising tool for designing the systems and therefore it was used for studying the process parameters for its interaction with the dyes and the affinity factors were compared for the systems with and without ligands. CIBACRON Blue F3GA and PROCION Red HE3G are the two affinity dyes used whereas PEG and Ammonium sulphate are the phase forming polymers used. Papain was purified using the optimal conditions which were obtained from response surface methodology. The resulting papain fractions were studied for purity in gel filtration chromatography.