Custom Protein Production and Expression in Biotechnology: Towards the Development of Biological Potential

To date in the biotechnology field, the synthesis of proteins to the customers’ specifications has had a drastic impact on the research and offered versatile opportunities in the applicability of proteins including in therapeutic applications and enzymatic engineering. Synthetic biology can be specifically defined as the production of a particular custom protein production by using engineered living systems to investigate the roles of proteins, the ways they interact, and the processes that generate such macromolecules or to use them for certain purposes.

Get to Know About Protein Production

Little did everyone know that they were part of a global protein production industry valued at more than seventy billion dollars. Proteins are amongst the most important biomolecules that have many tasks in the organisms’ including the participation in metabolism reactions, the provision of structure, the control of cellular signal transduction and the immune response. Every protein is composed of amino acids linking together in a chain, with this chain being made from a specific gene for the protein.

The generation of a particular custom protein expression entails incorporating the corresponding gene that encodes the protein, into a biological system to read the code and replicate the protein. This process has sequential steps which are gene cloning, protein expression, and lastly protein purification, all steps are important in determining the necessary yield and functionality of a protein.

Steps in Custom Protein Production

Cloning of Gene and Designing Vector

The first step in making proteins from any particular gene is to extract the gene in question. They are introduced into a circular piece of DNA referred to as a plasmid vector into which this gene is incorporated. The vector also possesses other regulatory sequences including promoters, terminators, and ribosome binding site – sequences that dictate how the gene will be expressed once it is placed in the host cell. In some cases, the scientists may prepare the gene sequence for expression in the system of choice through a process referred to as codon optimization.

Transformation into Host Cells

When the plasmid has been developed, it is then transferred to a host where the gene can be active. Depending on the requirements set by an experiment or an industrial process, the host system can be chosen. Host systems are generic nouns referring to bacteria, yeasts, insects’ cells, and mammalian cells- the most common host systems. The method of getting the plasmid into the host cells is different but one of the most preferred is drug or electrical means that forces the host cells to accept the plasmid and incorporate the new gene into their machinery.

Protein Expression

After infection with the virus-DNA, putting it in concrete terms, the host cells start manufacturing the required custom protein. The general regulation of protein synthesis can be constitutive where the protein is synthesized at all times or inducible where synthesis is dependent on certain chemicals or environmental stimuli. Since the protein synthesis machinery is highly sensitive to changing biochemical conditions of the cell, it is pivotal to maximize the chances of correct protein folding and subsequent protein functionalization by fine-tuning the expression conditions of the protein of interest, including temperature, nutrient availability, and temporal regulation.

Purification of Protein

Having been expressed the protein has to be purified from the host organism or from a culture medium. Purification of the protein is commonly done using several techniques composed of a step that calls for breaking down the host cells where the protein is found, other procedures include affinity chromatography, size exclusion, or ion exchange chromatography. The objective is to achieve an enriched, desired quality protein ideal for subsequent test or functioning.

Protein Characterization

The last step is, of course, to characterize the protein in order to determine the true identity and structure as well as its function. Biochemical techniques including mass spectrometry, SDS-PAGE and Western blot mineralize the size and purity of the protein. Also, counterpart functional assays are carried out to confirm how the protein will function when delivered to biological systems or into various industrial processes.

Expression Systems in Custom Protein Production

The selection of the appropriate expression system is one of the critical decisions that define the productivity, the quality of synthesized protein, as well as the possibility of its large-scale production. Below are some commonly used expression systems:

Bacterial Expression Systems

E.coli and its other kinds of bacteria are most frequently used in the production of protein since they grow much faster, can be genetically modified, and are relatively cheap. Bacterial systems can be very effective in generating large amounts of protein in a relatively short time. They are however less useful in the synthesis of proteins that need further modification after synthesis such as glycosylation that is found in other organisms of higher complexity.

Yeast Expression Systems

Bacterial systems include systems derived from bacteria and their genes like Escherichia coli, while mammalian systems involve systems derived from mammalian and their genes for instance, mice; yeast system is intermediate between bacterial and mammalian systems; for instance, yeast, Saccharomyces cerevisiae. Higher yeasts are comparatively easy to cultivate, capable to a certain extent of post-translational modification, and, therefore, these are more suitable for synthesizing eukaryotic proteins. Yeast systems are widely used in Industrial enzyme production and Vaccines production.

Insect Cell Expression Systems

Insect cells are very useful for synthesizing proteins that require proper folding and other co-translational modifications as seen in the Baculovirus expression system. This system gives higher levels of functional proteins in contrast to bacterial or yeast systems, which simplifies deposition of proteins appropriate for structural biology and matters constituent in production of vaccines.

Mammalian Cell Expression Systems

CHO cells are one of the most developed expression systems and are popular in the commercial production of therapeutic proteins including monoclonal antibodies and growth factors. The present study also demonstrated that recombinant mammalian cells are capable of performing the necessary post-translational modifications and synthesizing proteins that are very similar to human proteins. Although they are somewhat more difficult to cultivate and are much dearer in terms of costs and time.

Conclusion

Protein synthesis and production are indispensable events in contemporary biotechnology that continuously underpin medicine, industry, and research. Improvements in expression systems, gene editing, and protein refolding or purification methods further extend the prospects for custom protein synthesis to yield biological functions of proteins as desired for different uses.