Method for promoting secretory expression of rhIL-24 by engineering cell strain by using sodium butyrate

Abstract

The invention provides a method for promoting secretory expression of rhIL-24 by an engineering cell strain by using sodium butyrate, belonging to the technical field of gene expression. On the basisthat a site-specific integrated engineering cell strain FCHO / IL-24 for secretory expression of the rhIL-24 is subjected to adherent culture with a culture medium containing 10% of serum, adherent culture with a culture medium containing 0.5% of serum adherence and suspension culture with a culture medium containing 0.5% of serum, the cell strain is treated by using sodium butyrate with different final concentrations (0, 0.125, 0.25, 0.5, 1 and 2 mmol / L). According to the invention, NaBu can improve the secretory expression level of foreign protein by CHO cells, promotes the secretory expression level of rhIL-24 by engineering cells, promotes generation of G0 / G1 phase arrest of the engineering cells, and can change the metabolic activity of the cells under adherent culture conditions; and by using of the NaBu as an additive of a serum-free culture medium of the cell strain, the secretory expression level of the rhIL-24 by the cell strain is improved in high-density suspension culture, so experimental data is provided for large-scale culture of FCHO / IL-24 cells, and a reference can be provided for expressing gene engineering proteins by using mammalian cells at the same time.

Description

technical field [0001] The invention relates to the technical field of gene expression, in particular to a method for using sodium butyrate to promote the secretion and expression of rhIL-24 by engineered cell lines. Background technique [0002] Melanoma differentiation-related gene-7 / interleukin-24 (mda-7 / IL-24) encodes a protein consisting of 206 amino acid residues with a molecular weight of about 23.8kDa. IL-24 has broad-spectrum and specific anti-tumor activity, can specifically inhibit the growth of various tumors in vitro and in vivo, and selectively induce tumor cell apoptosis, with little effect on normal cells. At the same time, IL-24 has the ability to stimulate the body's anti-tumor immune response. It can not only stimulate peripheral blood mononuclear cells to secrete and produce inflammatory cytokines, such as IL-6, IL-1β, IL-12, GM-CSF and IFN- γ, etc., can also increase CD3 + CD8 + The number of T cells can trigger the body to produce an immune response ...

AI Technical Summary

Problems solved by technology

The activity of rhIL-24 expressed in mammalian cells is much higher than that expressed in bacteria, but the expression level is still extremely low

In the research on the expression of rhIL-24 in mammalian cells, there is no related technical research on the use of sodium butyrate to improve the expression of rhIL-24 in engineered cell lines

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