The analysis of recombinant insulin production technologies
DOI:
https://doi.org/10.24959/nphj.24.158Keywords:
insulin, recombinant DNA technology, expression system.Abstract
Aim. To analyze current trends in insulin production and expression systems used to obtain recombinant proteins.
Materials and methods. The specified goal was achieved using the methods of content analysis, comparative, logical, analytical and generalization of information. The research materials were publications in scientific periodicals, official websites of manufacturing companies.
Results. It has been found that the following expression systems are traditionally used for the commercial production of recombinant human insulins: bacterial – Escherichia coli, and yeast – Saccharomyces cerevisiae. But to meet the global need for insulin, it is necessary to develop new effective expression systems. Therefore, the use of other eukaryotic insulin expression systems that would be suitable for large-scale production is currently being studied. Among such systems, transgenic plants, which are characterized by the absence of potential human pathogens and the presence of post-translational protein modification mechanisms similar to human ones, are promising for introduction into insulin production. Currently, studies using Arabidopsis thaliana, tobacco, lettuce and strawberry plants are successful. Stem cells – embryonic, mesenchymal and induced pluripotent are also considered promising. The methods used in the large-scale production of recombinant insulin based on the synthesis of proinsulin and the two-chain method have been characterized. The technological flowcharts of both technologies have been drawn up, the advantages and disadvantages of each are given.
Conclusions. The analysis of data from the scientific literature has shown that the main methods of producing insulin preparations since the 1980s remain pro-insulin and two-chain methods, which are based on recombinant DNA technologies. The pro-insulin method is considered more efficient and cost-effective compared to the two-chain method since this technology consists of working with a single recombinant strain. The choice between these two methods depends on the specific needs of the manufacturer: for some cases a fast method with proinsulin synthesis is rational, for others – a more accurate and controlled two-chain method. The bacteria – Escherichia coli and yeast – Saccharomyces cerevisiae remain the expression systems used in the existing large-scale production technologies. But using E. coli, insulin precursors are produced in inclusion bodies, and fully functional polypeptides are obtained by solubilization and refolding steps. The yeast-based system yields a soluble insulin precursor that is secreted into the culture fluid, but requires prior humanization due to the risks of an immune response in humans. Thus, current production technologies cannot meet the growing demand for available insulin due to limited production capacity and high production costs, so research is being conducted to find new efficient expression systems, such as plant and mammalian cells, including Arabidopsis thaliana, tobacco, lettuce, strawberry, stem cells.
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