Production of Chloroplast-Derived Human Interferon Alpha

Introduction

Interferons (IFNs) appear after the body is infected with viruses and are involved in animal antiviral defense, cell growth regulation, and immune system activation. Interferon is used to treat a variety of diseases, including certain types of cancer and hepatitis C. Interferons alpha, beta and gamma are three naturally occurring interferons. Currently the only commercially produced interferon alpha (hIFN). hIFN is a broad range of bioactive cytokines with many therapeutic applications in the treatment of various human cancers and viral-derived diseases. However, the production of hIFN from microbial to mammalian expression systems has certain limitations in terms of cost, scalability, safety, and authenticity. IFNα 2b is widely used to treat hairy cell leukemia but is expensive. The development of modern biotechnology has enabled genetically modified crops to obtain large amounts of complex proteins in a cost-effective manner.

Fig. 1. IFN-2b signaling via JAK-STAT pathway. (Asmana Ningrum R, 2014)

Solutions

Our engineers are passionate about engineering chloroplasts to produce therapeutic proteins for research to treat or prevent human or animal disease. Lifeasible is an expert in the field of recombinant protein research and development, with a variety of mature and comprehensive platforms and technologies, providing global customers with professional solutions for the production of chloroplast-derived interferon alpha. Here, we provide a method for expressing recombinant IFNα 2b in transgenic tobacco chloroplasts using a polyhistidine tag. In addition, we performed chloroplast transformation to express interferon alpha 5 and 2 to facilitate integration of the transgene into the chloroplast genome by homologous recombination. Our solution process is roughly as follows:

(1) pTZ57R, pBCSK+ and pUC19 plasmid vectors are used to clone various fragments and construct chloroplast transformation vectors.
(2) The vector was transformed into tobacco chloroplasts by Agrobacterium transformation method.
(3) Transgenic tobacco with IFN-α2b sequence was screened by transient expression detection, and cultured and regenerated.
(4) Determination of interferon activity.
(5) Southern blot analysis whether the IFN-α2b gene is stably integrated into the tobacco chloroplast genome.
(6) Western blot analysis of the expression stability of IFN-α2b protein.

Features of Our Solutions

• IFN-α2b levels were as high as 20% of total soluble protein.
• Chloroplast-derived IFN-α2b has the ability to protect cells from viral infection.
• Chloroplast-derived IFN-α2b has antitumor and immunomodulatory properties.
• N. excelsior was chosen as the transient expression host.
• Chloroplasts can properly fold IFNα 2b and form appropriate disulfide bonds, reducing the cost of in vitro processing.

The chloroplast-transformed human interferon alpha is safe and effective, and also treats hepatitis and cancer caused by viral infections. Lifeasible has extensive knowledge and experience in the development and production processes of the chloroplast-derived human interferon alpha. Our mission is to provide customers with comprehensive, reliable, professional solutions to accelerate your research. If you are interested in our solutions for the production of chloroplast-derived human interferon alpha, please contact us at any time.

References

1. Nurjis F. (2010) Chloroplast-based overexpression of Pharmaceuticals to develop Cost-effective Therapeutics[D]. Quaid-i-Azam University Islamabad, Pakistan.
2. Asmana Ningrum R. (2014) Human interferon alpha-2b: a therapeutic protein for cancer treatment[J]. Scientifica.