The purification of recombinant proteins is a long and complex process, as our protein purification handbook proves, and is influenced by a multitude of variables. As has already been mentioned, the nature of the protein itself, its properties and the characteristics of the host producer will greatly influence the design and development of this procedure. There are also very few available techniques, although it is very well known that the preferred one is affinity chromatography.
During the sample preparation for the purification of recombinant proteins, there are several considerations that will influence its development. All preparation steps, which are thoroughly explained in our protein purification handbook, are important and we have to pay attention to them.
Chromatography is one of the most common methods for the purification of recombinant proteins, and more specifically affinity chromatography is the one that is mostly used due to its high specificity, which allows us to obtain great purity in one single step. When using this technique, a tag is added to the protein of interest, a small structure that is not included in the original protein and that allows us to easily capture it. As we explain in our protein purification handbook, tags are generally short sequences of 3-4 amino acids (up to a maximum of 15) and are intended to minimize as much as possible the properties of the protein.
Recombinant proteins are produced in the host cells along with a great variety of molecules that it contains naturally. However, for most of their applications (such as for example in therapeutics), a recombinant protein should be purified and isolated from the rest of cell molecules. As we explain in our protein purification handbook, this is obtained by the recombinant protein purification processes.
The host in which a recombinant protein is produced doesn’t naturally include the gene of this protein in its genome. Therefore, this gene needs to be introduced in a process called molecular cloning. Successful cloning of a gene requires several elements, which are discussed in our protein purification handbook.
As you can learn in our protein purification handbook, production techniques of recombinant proteins offer multiple options when it comes to available hosts; in other words, the organisms that incorporate the gene of the protein of interest and express it correctly.
As you will learn in our protein purification handbook, proteins are biomolecules of great value to humans, since they have a wide variety of uses in different sectors. They can have therapeutic (such as in insulin cases or blood clotting factors), industrial (such as the lipases that are included in detergents that degrade grease stains) or biotechnological (in the case of certain toxins that are used as pesticides) applications. They also have an extensive application in the research field, in cases when they can be used to study the molecular mechanisms of many diseases, find new drugs, or elucidate tridimensional structures, among others.
Human beings extensively use proteins for different purposes such as health, industrial production or food. Nevertheless, most of these proteins come from natural sources that do not produce them in sufficient quantities for human use; therefore, we turn to the use of recombinant proteins. Such proteins are obtained from organisms (called “host organisms”) that do not produce them naturally, and in which the gene of the protein of interest is incorporated.
