Expression of Difficult-to-Express Proteins, Human IL-12 and IFN-<i>γ</i>

Abstract

It is known to be that lactic acid bacteria induce the IL-12. IL-12 activates NK cells and promotes the production of IFN-γ. IFN-γ activates macrophages, resulting in enhanced phagocytosis and bactericidal activity. We have been investigating fermented foods that activate the immune function. For that purpose, a specific antibody is required. We tried to express IL-12p35 by the usual method, but IL-12p35 was not expressed at all. In the present study, we constructed, purified human IL-12p35 and obtained a specific antibody against IL-12p35. We also purified human IFN-γ and obtained specific antibody against IFN-γ. We have established a method for expressing poorly expressed proteins. The method we have established can be applied to the purification of poorly expressed proteins and antibody production.

Fermented food Tempeh induces interleukin 12 and enhances macrophage phagocytosis

“…29-37. https:// www.scirp.org/journ al/paper infor mation.aspx?paper id=107559). *p < .05, compared with PBS IL-12p35, and immunized the protein to rabbit (Hosaka et al, 2021).…”

“…Samples were analyzed by SDS/PAGE, followed by immunoblotting. The PVDF membrane was reacted with an IL-12p35 antibody (Hosaka et al, 2021) (diluted 500 times) with an anti-rabbit AP IgG (BioRad, diluted 10,000 times) or an anti-rabbit HRP IgG (Biorad, diluted 15,000 times). Samples were treated with BCIP-NBT solution (Nacalai Tesque) or an ECL Plus Western Blotting Detection System (Cytiva).…”

“…We constructed, then purified human IL-12p35 and IFNγ using the E. coli expression system (Hosaka et al, 2021). We also immunized the purified proteins to rabbit and obtained antiserum against IL-12p35 and IFNγ.…”