Enzyme Purification英文电子书.pdfVIP

Protein purification What is protein purification? What is protein purification? is extraction of an almost single enzyme or protein from cells is extraction of an almost single enzyme or protein from cells , tissue, bacteria…etc, which may contain more than , tissue, bacteria…etc, which may contain more than What is the need for a pure protein? What is the need for a pure protein? Three Phase Purification Strategy Three Phase Purification Strategy Capture phase: Capture phase: Isolate, concentrate and stabilise the target product. The product should be Isolate, concentrate and stabilise the target product. The product should be concentrated and transferred to an environment which will conserve potency concentrated and transferred to an environment which will conserve potency /activity. At best, significant removal of other critical contaminants can also be /activity. At best, significant removal of other critical contaminants can also be achieved. achieved. Intermediate purification phase: Intermediate purification phase: the objective is to remove most of the bulk impurities such as other proteins and the objective is to remove most of the bulk impurities such as other proteins and nucleic acids, endotoxins and viruses. nucleic acids, endotoxins and viruses. Polishing phase: Polishing phase: Achieve high purity by removing any remaining trace impurities or closely related Achieve high purity by removing any remaining trace impurities or closely related substances substances The physical boundaries of protein stability: The physical boundaries of protein stability: -pH: determine pH where enzyme is most stable