Protein optimization is one of the challenging aspects. According to researchers, obtaining large quantities of active protein is the greatest challenge. Optimization of protein production constantly depends on significant factors such as culture conditions, purification, and expression construct creation.
However, most biomedical specialists have been scientifically committed to analyzing the various methods to optimize protein expression and production.
In essence, biomedical specialists are more concerned with the expression and production of recombinant proteins.
Although the purification and production of recombinant protein are available in various ways, most specialists are committed to identifying artificial and traditional methods that can help in protein production optimization.
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The following article will provide in-depth information on how to optimize protein production.
Choosing an appropriate expression host
To optimize protein production, you must first choose the appropriate expression host, such as the Escherichia coli, insect cells, and mammalian cells.
According to scientific statistics, most proteins prove to be fully expressed in Escherichia coli. Therefore, the choice of the expression host will determine and affect the quantity production of protein.
The protein expression on Escherichia coli contributes to over 61% protein production compared to the other host. Escherichia coli can be categorized as the most convenient and most used recombinant expression host to optimally produce protein due to their inexpensive features, speed, and easy scale-ups.
However, choosing Escherichia coli cells as the expression host will require an additional choice of the appropriate purification tools and protocols.
There is also a high increase in the use of eukaryotic hosts in protein expression, which has contributed more to the rise in the production rate of membrane protein.
Protein from hosts such as mammalian cells proves to be challenging to express and optimize. The use of hot expression cells is one of the discovered engineering ways to effectively ensure protein optimization as it helps in overcoming the possible bottlenecks in the production process.
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Selecting the type of promoter
To optimize protein production, you have to choose the right type of promoter to use. Selecting the best and strongest promoter will guarantee the production of large amounts of protein.
Since the most used expression host in protein production in Escherichia coli, T7 is the preferred promoter found in the expression host.
Other promoters include polyhedron, which is found in the baculovirus expression host, and the CMV promoter found in the mammalian cells.
However, if you choose a strong promoter, you can boost its strength to increase the production rate.
To increase protein production rate, you can reduce the transcription rate by introducing a more potent promoter or a point mutation. Alternatively, you can quickly reduce the level of polymerase in the host cell.
Gene translation is also a determining factor in optimizing protein production. The introduction of viruses in the cells may lead to mutilation of the genes hence causing translation and thus leading to a decrease in protein expression rate.
For example, antibody production may be affected by the introduction of antigens which alters the B-lymphocytes, which are responsible for producing the antibodies.
Another example is how the Ebola virus affects translational silencing, leading to increased recombinant protein production.
Selecting host cell
Though you can choose a more potent promoter, the choice of the host cell will also affect protein production. For example, a poor promoter silencing on the host cells will negatively affect the quantity and production rate.
When a mammalian cell is used, poor expression and epigenetic of the CMV promoter could lead to a low production rate.
However, this can be corrected through the nuclear attachment region or by including the initiation region, which will help in improving recombinant protein production in the mammalian cell lines.
The last significant step to optimize protein production is choosing the appropriate purification method. The purification process of the recombinant protein can be long and complex.
The process is majorly affected by a variety of factors. Therefore, choosing a suitable purification method will affect protein quantity and production rate.
For example, Escherichia coli is one of the most used expression host systems to optimize protein production. Therefore, for the purification process, there are tools and products that you can choose to ensure optimum presentation of protein—for example, plasmid, resin system, strain type, and affinity tag.
Besides, the choice of the purification product or tool will affect the protein production rate. The main goal in the protein production process is to separate the recombinant protein from the stabilization product.
Therefore, a high concentration level is needed to collect large volumes of recombinant protein. Choosing a suitable purification method will guarantee optimization of protein production and the end of the process.
In conclusion, protein production, expression, and purification are determined by many other factors that this article may not have tackled. However, the above steps are crucial in ensuring the optimization of protein production. During the production process, you have to consider providing an optimum temperature conducive to the survival of the proteins.