白介素24融合蛋白的表达、纯化和促伤口愈合功能的鉴定

本研究通过优化小鼠白介素24(mIL-24)基因序列,构建以Fc-tag为标签的mIL-24-Fc融合蛋白.对CHO进行MTX分级加压筛选,获得了稳定、高表达mIL-24-Fc蛋白的阳性克隆,同时优化了CHO细胞无血清驯化与稳定表达蛋白体系.经protein A亲和层析纯化mIL-24-Fc蛋白并利用AP binding实验检测生物学活性后,小鼠皮内注射mIL-24-Fc蛋白和伤口愈合实验证明:注射mIL-24-Fc蛋白后,小鼠表皮产生2~3层细胞增生,伤口愈合速度加快.

Expression, purification and identification of wound healing function of Interleukin-24 fusion protein

Interleukin-10 family is one of the important cytokine that can cause psoriasis related diseases, and interleukin-24 is relatively important in cascade effect. As a new discovery member of interleukin-10 family, we aim to study the role of interleukin-24 in psoriasis and pro-inflammatory response. In this study, we use biosynthetic gene interleukin 24 mice (mIL-24) to built mIL-24-Fc fusion protein. Then the recombinant plasmid was transfected into CHO cell. After serum-free culture in CHO, we purified mIL-24-Fc fusion protein by protein A affinity chromatography and confirmed by AP binding assay. The result showed that after intradermal injected mice, a large number of keratinocytes showed two to three proliferation and in wound-healing process, mIL-24-Fc can promote epidermal hyperplasia and accelerate wound healing. These findings mean that interleukin424 plays an important role in the inflammatory response, and medical value.