Enhancing the Expression and Purification of IL-29: A study of autoinduction and one-step Purification Methods

Zia Ur Rahman, Nadeem Ahmed, Numan Fazal, Muhammad Islam Khan, Mohsin Ahmed Khan, Saad Tahir, Muhammad Akram, Sajjad Ullah, Ahmad Usman Zafar


Background: Cytokines have long been viewed as a sign of hope due to their immunomodulatory and therapeutic characteristics. Developing simple, economical and readily scaled technologies to simplify their manufacturing is a critical challenge.

Method: In this study we have used a customized medium to automatically induce the expression of the IL-29 in E. coli expression system from the T7 promoter, allowing for higher yields as compared to the traditional technique of IPTG induction. Similarly, one-step purification method is employed to make the fermentation process cost-effective, along with enhancing its efficiency.

Results: From 1 L batches of IPTG-induced and autoinduced media, the harvested biomass was 11.8 g and 13.4 g, respectively and their corresponding IBs were 3.8 g and 4.8 g. Total protein purified from 1 L batch was 132  mg, at a concentration of 13 mg/mL, with an indicated high purity of 97%. IL-29 significantly decrease the metabolic activity of HepG2 cells. Specifically, 50% of the cells died at a concentration of 0.156 μg/mL, while 80% of the cells died at a concentration of 5 μg/mL.

Conclusion: This study presents an economical solution for producing and purifying IL-29 in E. coli, resulting in higher yields of biomass and IBs than expensive traditional method. The purified protein was highly pure and had immunomodulatory effects on HepG2 cells. These findings have important implications for developing simplified and scalable technologies for cytokine production with therapeutic potential.

Keywords: Escherichia coli; Cytokines; Interleukins; Interferons; Protein purification     

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