What are the principles of protein purification

SCG Protein Purification System Co., Ltd.-Spectro Instruments has organized the contents of protein purification principles for everyone.

For protein purification, the inherent similarities and differences between different proteins should be used, and the similarities between various proteins should be used to remove the contamination of non-protein substances, and the differences in each protein should be used to purify the target protein from other proteins. The size, shape, charge, hydrophobicity, solubility and biological activity of each protein will be different, and these differences can be used to extract proteins from mixtures such as E. coli lysates to obtain recombinant proteins.

Protein purification is roughly divided into two stages: crude separation stage and fine purification stage. The general method of protein purification is the resin method. The crude separation stage mainly separates the target protein from other cellular components such as DNA and RNA. Due to the large sample volume and complex components at this time, the resin used requires high capacity, high flow rate, large particles, and wide particle size distribution. And can quickly separate the protein from the contaminants, if necessary, can participate in corresponding protective agents (such as protease inhibitors) to prevent the target protein from being degraded.

The fine purification stage requires higher resolution. This stage is to separate the protein of interest from those protein regions with close molecular weight and physical and chemical properties. Smaller resin particles are used to improve resolution. Ion exchange columns and hydrophobic For column application, the two factors of resin selectivity and column efficiency should be considered.

Selectivity refers to the specificity of resin binding to the target protein, and column efficiency refers to the ability of each protein component to elute from the resin one by one. The narrower the elution peak, the better the column efficiency. With only good selectivity, the elution peak is too wide, and the protein cannot be separated as usual.

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