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Protein Purification Part 11

Tham khảo tài liệu 'protein purification part 11', khoa học tự nhiên, công nghệ sinh học phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | The Denaturing and Renaturing are Critical Steps in the Purification of Recombinant Protein in Prokaryotic System 143 Fig. 3. Purification of rhIL-lRaST by S-Sepharose According to UV absorption peak fractions 6 7 8 9 and 10 were collected and determined for its rhIL-1Ra concentration and purity 144 Protein Purification Fig. 4. Picture of Commassie-Blue staining after SDS-Page of purified rhIL-1Ra Lane 0 Markers 1 Flow through solution of rhIL-1RaST after DEAE-Sepharose purification 2 rhIL-1Ra before S-Sepharose purification 3 rhIL-1Ra after S-Sepharose purification 4.2 The denaturing and renaturing of recombinant protein in inclusion bodies If the protein exists in inclusion bodies form denaturing and renaturung treatments are mandatory for the purification of this protein. An ideal denaturing and renaturing system must serve the following purpose high-output low-loss and normal bio-activity final products. In industrial level the ideal renaturing technology must also be easily amplified to produce large-scale products. Must commonly used denaturing solution is 6 M guanidine chloride or 8 M urea. There are some renaturing methods including dialysis dilution ultra-filtration. Some additives can be applied into the renaturing system to enhance the refolding efficacy such as denaturing agents PEG detergents redox system L-Arg chaperone etc. However there are no general guidelines that can be applied to all refolding systems and one must test the efficiency of each method to ensure the optimized output. Some commercial available kits can be used to test the optimized renaturing system. Here are a few samples of protein expressed in inclusion bodies Monokine induced by IFN-y Mig is a CXC-chemokine CXCL9 . It plays important roles in regulation of immune activities and knowledge of the protein in areas of allograft transplants autoimmune diseases and cancer therapy is evolving quickly. Mature murine MIG is a protein of 106 amino acids with molecular weight of 14.4 kDa.