Precipitation by Change in PH or by Adding a Chelating agent, Crude homogenate, Bovine pancreas, Polymer ligand, Copolymerization, N-acetyl glucosamine

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Precipitation by change in pH or by adding a chelating agent. Following are some examples of the use of affinity precipitation for Bioseparation and purification by altering pH or by adding a chelating agent: (i) Purification of trypsin from a crude homogenate of bovine pancreas was achieved using a polymer ligand, which was synthesized by copolymerization of N-acryloyl-m-amino-benzamidine and acrylamide. The polymer with trypsin could be precipitated below pH 4.0 and redissolved above pH 8.0. (ii) Similarly, wheat germ lectin was precipitated using chitosan (a polymer of partially deacetylated N-acetyl glucosamine), which precipitates at pH 5.5 and redissolved by a shift in pH. (iii) Lysozyme could also be precipitated using chitosan could also be precipitated using chitosan at pH 4.0. (iv) Lectins from rice, potato and tomato have been precipitated using N-acetylglucosomine and chitosan as ligands.

(v) Human alkaline phosphates was purified using concanavalin A as the ligand. (vi) Alginate, which can be precipitated by addition of ca²⁺ ions has found wide spread application in affinity purification of proteins and enzymes like α-amylases, phospholipases and polygalacturonases, which are used in food industry. The bound enzyme can be eluted with maltase and reversible solubilization can also be achieved by adding a chelating agent like EDTA. (vii) Galactomannon is another compound which could be coupled with protein A and used for precipitating human IgG. Methacrylic acid, methyl methacrylate, eudragit and microbial lipase are other ligands, which have been used for bioseparation and purification.

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Indirect Methods for Monitoring Cell Concentration Disintegration of Cells Separations Concentration and Purification of the Product Affinity Precipitation Precipitation by Change in pH or by Adding a Chelating Agent Affinity Thermoprecipitation Immobilized Metal ion Affinity Chromatography IMAC Expanded Bed Chromatography Drying of the Product Downstream Processing in India Downstream Processing Separation of Cells/Biomass