Prospects of protein separation and purification methods
In recent years, with the rapid development of various disciplines, the demand for protein purification technology has continued to increase, existing purification methods have been increasingly improved, and new purification methods have also emerged one after another. Hydroxyapatite is the crystal of calcium phosphate, due to its physical and chemical properties It is not stable enough, the binding ability is poor, and it is difficult to use for chromatography. Coming in Bio-Rad has improved it and increased the ratio of calcium and phosphorus to form spherical, porous, and stable ceramic hydroxyapatite particles. Positively charged calcium ions and negatively charged phosphate ions can be combined with the carboxyl and amino groups of the protein. By adjusting the pH of the buffer, acidic and basic amino acids can be selectively combined with this resin to change the salt of the buffer The concentration can elute and separate the protein.
In terms of affinity purification, Sigma has developed a purification method using FLAG tags. FLAG has a small molecular weight and hydrophilicity. The protein expressed by fusion with FLAG is not easy to form inclusion bodies and its activity is not affected. Other new tags and corresponding purification systems also have CBP, BCCP tag and avidin-biotin purification system, maltose binding protein tag, etc. I believe that as time goes on, there will be more and better methods to fully apply these methods to toilet research, biopharmaceuticals, vaccines, diagnostic reagents Development and other work will be of great help.
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