NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, China CDC, Beijing, 100052, China.
Beijing Furen Ruihui Biomedical Research Institute Co.,Ltd, Beijing, 100176, China.
BMC Biotechnol. 2020 Mar 13;20(1):16. doi: 10.1186/s12896-020-00605-2.
The type I human interferon (IFN) family consists of a group of cytokines with a multiplicity of biological activities, including antiviral, antitumor, and immunomodulatory effects. However, because the half-life of IFN is short, its clinical application is limited. Increasing the yield and biological activity of IFN while extending its half-life is currently the focus of IFN research.
Two novel long-acting recombinant human IFN-α2b (rhIFN-α2b) proteins were designed in which the carboxyl-terminal peptide (CTP) of the human chorionic gonadotropin β su bunit and N-linked glycosylation sequences were linked to rhIFN-α2b. They were designated IFN-1CTPON (fused at the C-terminus of rhIFN-α2b) and IFN-2CTPON (fused at both the C-terminus and N-terminus of rhIFN-α2b). Monoclonal CHO cell strains stably and efficiently expressing the IFNs were successfully selected with methotrexate (MTX), and the highest expression levels were 1468 mg/l and 1196 mg/l for IFN-1CTPON and IFN-2CTPON, respectively. The proteins were purified with affinity chromatography and molecular sieve chromatography. IFN-1CTPON and IFN-2CTPON showed antiviral and antiproliferative activities in vitro. Notably, the half-life of IFN-1CTPON and IFN-2CTPON in vivo were three-fold and two-fold longer than that of commercially available rhIFN-α2b.
CHO cell strains stably expressing long-acting rhIFN-α2b were screened. The purified IFN-CTPON protein has biological activity and an extended half-life, and therefore potential applications.
I 型人干扰素（IFN）家族由一组具有多种生物学活性的细胞因子组成，包括抗病毒、抗肿瘤和免疫调节作用。然而，由于 IFN 的半衰期短，其临床应用受到限制。提高 IFN 的产量和生物活性，同时延长其半衰期，是目前 IFN 研究的重点。
设计了两种新型长效重组人干扰素-α2b（rhIFN-α2b）蛋白，在其中连接人绒毛膜促性腺激素β亚基的羧基末端肽（CTP）和 N 连接糖基化序列。它们被命名为 IFN-1CTPON（rhIFN-α2b 的 C 末端融合）和 IFN-2CTPON（rhIFN-α2b 的 C 末端和 N 末端融合）。用甲氨蝶呤（MTX）成功地选择了稳定且高效表达 IFNs 的单克隆 CHO 细胞株，IFN-1CTPON 和 IFN-2CTPON 的最高表达水平分别为 1468mg/L 和 1196mg/L。用亲和层析和分子筛层析对蛋白质进行纯化。IFN-1CTPON 和 IFN-2CTPON 在体外具有抗病毒和抗增殖活性。值得注意的是，IFN-1CTPON 和 IFN-2CTPON 的体内半衰期分别比市售 rhIFN-α2b 长 3 倍和 2 倍。
筛选出稳定表达长效 rhIFN-α2b 的 CHO 细胞株。纯化的 IFN-CTPON 蛋白具有生物活性和延长的半衰期，因此具有潜在的应用前景。
