A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-alpha from Raphanus sativus L. Plants

Human interferon (IFN) is a type of cytokine that regulates the immune system's response to viral and bacterial infections. Recombinant IFN-alpha has been approved for use in the treatment of a variety of viral infections as well as an anticancer medication for various forms of leukemia. The objective of the current study is to produce a functionally active recombinant human IFN-alpha 2a from transgenic Raphanus sativus L. plants. Therefore, a binary plant expression construct containing the IFN-alpha 2a gene coding sequence, under the regulation of the cauliflower mosaic virus 35SS promoter, was established. Agrobacterium-mediated floral dip transformation was used to introduce the IFN-alpha 2a expression cassette into the nuclear genome of red and white rooted Raphanus sativus L. plants. From each genotype, three independent transgenic lines were established. The anticancer and antiviral activities of the partially purified recombinant IFN-alpha 2a proteins were examined. The isolated IFN-alpha 2a has been demonstrated to inhibit the spread of the Vesicular Stomatitis Virus (VSV). In addition, cytotoxicity and cell apoptosis assays against Hep-G2 cells (Human Hepatocellular Carcinoma) show the efficacy of the generated IFN-alpha 2a as an anticancer agent. In comparison to bacterial, yeast, and animal cell culture systems, the overall observed results demonstrated the efficacy of using Raphanus sativus L. plants as a safe, cost-effective, and easy-to-use expression system for generating active human IFN-alpha 2a.