Interleukin-30 (IL-30), or IL-27p28, is the α subunit of IL-27 constructed by Epstein-Barr virus induced gene 3 (EBI3) and IL-27p28 binding via non-covalent bonds. IL-30 can be independently secreted and function independent of IL-27. Recent studies demonstrated IL-30 could concurrently antagonize T helper 1 (Th1) and Th17 responses and might have therapeutic implications for controlling autoimmune diseases. However no reports have stated an efficient method to generate relatively large quantity of IL-30. In the present study, anE.coli expression system for the rapid expression of the mouse IL-30 is developed. For the first time, IL-30 was expressed in a form of soluble fusion protein and purified using a... More
Interleukin-30 (IL-30), or IL-27p28, is the α subunit of IL-27 constructed by Epstein-Barr virus induced gene 3 (EBI3) and IL-27p28 binding via non-covalent bonds. IL-30 can be independently secreted and function independent of IL-27. Recent studies demonstrated IL-30 could concurrently antagonize T helper 1 (Th1) and Th17 responses and might have therapeutic implications for controlling autoimmune diseases. However no reports have stated an efficient method to generate relatively large quantity of IL-30. In the present study, anE.coli expression system for the rapid expression of the mouse IL-30 is developed. For the first time, IL-30 was expressed in a form of soluble fusion protein and purified using a way of simple affinity chromatography. In order to avoid the impact of minor codons on expressing eukaryotic protein in E.coli and improve the expression quantity, the nucleotide sequence of IL-30 was optimized. The optimized gene sequence was then sub-cloned into the pET-44a (+) vector, which allowed expression of IL-30 with a fusion tag - NusA. The vector was transformed into E.coli and the expressed fusion protein, NusA-IL-30, was purified by Ni-chromatography. Then the fusion tag was removed by cleavage with thrombin. The purify of purified IL-30 was identified using SDS-PAGE as well as HPLC and the purify was up to about 92%. And the yield of IL-30 was 8.95 mg from 1 L of bacterial culture. Western blot confirmed the identity of the purified protein. The recombinant IL-30 showed its biological activity by inhibiting Th17 differentiating from Naïve CD4+ T cells. Therefore, this method of express and purify IL-30 provide novel procedures to facilitate structural and functions studies of IL-30.
