金属离子螯合色谱.pptx

Metal Chelate Affinity Chromatography;Protein purification?is a series of processes intended to isolate a single type of?protein from a complex mixture. Protein purification is vital for the characterization of the function, structure and interactions of the protein of interest.;Proteins are purified using chromatographic purification techniques which separate according to differences in specific properties;Affinity chromatography separates proteins on the basis of a reversible interaction between a protein (or group of proteins) and a specific ligand coupled to a chromatography matrix. ;Metal-Chelate Affinity Chromatography;;His-Tag for Purification of Recombinant Proteins;Key Parameters for the Operation of MCAC;Theory Model;Applications;Advantages;Unique characteristics IMAC chromatography Often allows single‐step purification procedures Allowing to investigate how the different metal‐ions affect the adsorption process without changing the matrix Has high protein loading capacities if compared to other affinity chromatographic techniques Is useful for concentrating dilute protein solutions Is compatible with a number of buffers containing high ionic strength or chaotropic components Generally does not affect the structure of proteins The use of a non‐charged IMAC column allows solutions to become transiently sterile since all metal‐ions essential for bacterial growth are removed by chelation ;Disadvantages;