Abstract
Biopharmaceutical production is currently a multibillion-dollar industry with high growth perspectives. The research and development of biologically sourced pharmaceuticals are extremely important and a reality in our current healthcare system. Interferon alpha consensus (cIFN) is a non-natural synthetic antiviral molecule that comprises all the most prevalent amino acids of IFN-α into one consensus protein sequence. For clinical use, cIFN is produced in E. coli in the form of inclusion bodies. Here, we describe the use of two solubility tags (Fh8 and DsbC) to improve soluble cIFN production. Furthermore, we analyzed cIFN production in different culture media and temperatures in order to improve biopharmaceutical production. Our results demonstrate that Fh8-cIFN yield was improved when bacteria were cultivated in autoinduction culture medium at 30 °C. After hydrolysis, the recovery of soluble untagged cIFN was 58% from purified Fh8-cIFN molecule, fourfold higher when compared to cIFN recovered from the DsbC-cIFN, which achieved 14% recovery. The biological activity of cIFN was tested on in vitro model of antiviral effect against Zika, Mayaro, Chikungunya and SARS-CoV-2 virus infection in susceptible VERO cells. We show, for the first time, that cIFN has a potent activity against these viruses, being very low amounts of the molecule sufficient to inhibit virus multiplication. Thus, this molecule could be used in a clinical approach to treat Arboviruses and SARS-CoV-2.
Highlights
Due to many scientific advances, with the use of modern molecular biology tools, the biopharmaceutical industry has become a multibillion-dollar industry
The production of cIFN fused to the Fh8 tag (HF-IFN) favored protein solubility when compared to the H- and HD-IFN fusion molecules
E. coli BL21(DE3) cultivation for production of HF-IFN in autoinduction medium at 30 ◦C presented a final recovery of soluble cIFN of 8 mg/L, 5.5-fold higher when compared to HD-IFN
Summary
Due to many scientific advances, with the use of modern molecular biology tools, the biopharmaceutical industry has become a multibillion-dollar industry. The biotechnology industry invested massively in the production of these molecules indicating a substantial growth dynamic in the future. The “Global Protein Therapeutics Market Outlook 2020” report estimates that by the end of 2020, this market may reach $208 billion dollars [1]. Biopharmaceuticals are characterized as molecules with therapeutic proprieties, produced through biotechnological processes. These drugs are currently used in a wide array of medical specialties and sometimes are considered the most effective treatments. Therapeutic proteins are used in a broad range of diseases, such as cancer, metabolic disorders and immunotherapy [2]
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