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.

中文翻译：

---

长效重组人干扰素-α2b的研制及其生物活性


I 型人干扰素 (IFN) 家族由一组具有多种生物活性的细胞因子组成，包括抗病毒、抗肿瘤和免疫调节作用。但由于IFN的半衰期较短，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 IFN-α2b (rhIFN-α2b) 蛋白，其中人绒毛膜促性腺激素 β 亚基的羧基末端肽 (CTP) 和 N 连接糖基化序列与 rhIFN-α2b 连接。它们被命名为IFN-1CTPON（融合在rhIFN-α2b的C末端）和IFN-2CTPON（融合在rhIFN-α2b的C末端和N末端）。利用甲氨蝶呤（MTX）成功筛选出稳定高效表达IFN的单克隆CHO细胞株，IFN-1CTPON和IFN-2CTPON的最高表达水平分别为1468 mg/l和1196 mg/l。用亲和层析和分子筛层析纯化蛋白质。 IFN-1CTPON和IFN-2CTPON在体外表现出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的rhIFN-α2b长三倍和两倍。筛选稳定表达长效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。