Essay on Applications of Protein Purification

Essay on Applications of Protein Purification

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Protein purification has a variety of applications in drug delivery, tissue engineering, and biointerface science. Elastin-like polypeptides can undergo protein purification so that a single type of protein can be isolated from a complex mixture. Because of the increased demands of peptide-based biomaterials for drug delivery and tissue engineering, efficient protein purification techniques must be used. Four techniques that will be discussed in this report are: Affinity chromatography, Centrifugation-based Inverse Transition Cycling (ITC), Microfiltration-based Inverse Transition Cycling, and finally protein purification by self-cleaving purification tags. The effect of different salts on the level of purity achieved will also be discussed.


The first and the most popular method to purify proteins is Affinity Chromatography, which is based upon molecular conformation1,2,3. This method uses a chromatography column that contains packing materials (resins). These resins have ligands attached to their surfaces, which allows them to recognize and bind to the protein of interest, and hence easily separate the particular compounds. Chromatography separates proteins based upon their size, charge, hydrophobicity, and shape. A single chromatographic step or multiple chromatographic steps (Ex: size followed by shape) can be applied to achieve the desired purity. While this scheme works well for small-scale purification, it is generally not recommended to use this technique for purification of large quantities of elastin-like polypeptides because of the high cost of affinity chromatography resins and the affinity columns. The column packing materials can be reused after regeneration; however, the performance of the chromatography colum...


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...ion steps, making it possible to conduct these steps at room temperature. Hence, this procedure was more energy efficient, with a gentler purification method. Therefore, purification of ELP by salt substitution eliminates affinity resins and their columns, which lowers the cost of purification, resulting in a more economical method.


The readily available need for purified proteins is important for many biomedical applications such as therapeutics and diagnostics, and in regenerative medicine and biosensing. Also, these purified proteins are an important factor in drug discovery. By recombinant expression, many proteins can be produced in larger quantities. However, the cost of the final product (~70%) is due to the cost of purification. Over the years, many strategies have been employed and are still being researched in order to improve protein purification.

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