Production of Chloroplast-Derived Human Interferon Gamma

Introduction

Interferon gamma (IFNγ) is a therapeutic cytokine that plays other roles in the immune regulation of pathogenic bacterial and viral infections and is also used to prevent viral proliferation. To date, it has been approved for the treatment of chronic granulomatous disease and malignant osteosclerosis. Researchers have engineered interferon gamma in a number of different expression systems, and the resulting recombinant human interferon gamma is a lucrative biopharmaceutical. The product of human interferon γ expressed in prokaryotic expression system is not glycosylated and the purification process is cumbersome, which is limited. Other expression systems have also not shown satisfactory results in terms of yield, biological activity or economic viability of the protein. The possibility of chloroplast transformation in higher plants increases the appeal of this expression system for the production of pharmaceutical proteins at biomanufacturing sites. Several mammalian-derived proteins have been successfully expressed in tobacco chloroplasts, such as human growth hormone, high human serum albumin, interferon alpha2b, insulin, etc.

Fig. 1. Expression of recombinant human interferon gamma in various expression systems. (Razaghi A, et al., 2014)

Solutions

Our engineers are passionate about engineering chloroplasts to produce therapeutic proteins for research to treat or prevent disease in humans or animals. 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 gamma.

The application of interferon gamma as an anticancer immunotherapeutic agent is increasingly promising. Our engineers attempted to develop a low-cost plant system to produce transgenic plants containing the interferon gamma gene (ifnG), such as rice, tomato, tobacco and rapeseed. Here, we provide a method for expressing recombinant interferon gamma gene in tobacco chloroplasts using aadA marker. In addition, we will also verify the feasibility of dual construct expression of ifnG and marker gene (aadA) in one operon. Our solution process is roughly as follows:

(1) IfnG was inserted into the plastid transformation vector pKCZ between Prrn and RBS of aadA.
(2) We transformed the above vectors into tobacco chloroplasts using a gene gun-mediated method.
(3) Whether the IfnG gene was stably integrated into the tobacco chloroplast genome and homogeneity were analyzed by polymerase chain reaction (PCR) and Southern blotting.
(4) Western blot analysis of IfnG protein expression in regenerated plants.
(5) Quantitative analysis of the expressed IFNg by enzyme-linked immunosorbent assay (ELISA).

Features of Our Solutions

• Bicistronic expression of ifnG and aadA genes in tobacco chloroplasts simplifies genetic manipulation.
• Tobacco plastid transformation is an efficient tool for large-scale production of recombinant interferon gamma protein.
• The INF-γ produced by transgenic chloroplasts can reach 3% tsp.
• Interferon-gamma has a short half-life of 4-6 hours.
• The IFN-γ produced by the transgenic chloroplasts has the same biological activity as the natural human IFN-γ.

The chloroplast-transformed human interferon gamma is safe and effective, and also treats chronic granulomatous disease and malignant osteosclerosis. Lifeasible has extensive knowledge and experience in the development and production processes of the chloroplast-derived human interferon gamma. 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 gamma, please contact us at any time.

References

1. Razaghi A, Owens L, Heimann K. (2016) Review of the recombinant human interferon gamma as an immunotherapeutic: Impacts of production platforms and glycosylation[J]. Journal of biotechnology. 240: 48-60.
2. Razmi S, Javaran M J, Bagheri A, et al. (2019) Expression of human interferon gamma in tobacco chloroplasts[J]. Rom. Biotechnol. Lett.. 24: 208-215.