IFN-λ1 in CHO cells: its expression and biological activity

• Published in: Cellular & molecular biology letters 2017-12, Vol.22 (1), p.26-26, Article 26
• Main Authors: Yuan, Wu-Mei, Zhang, Wan-Ju, Ma, Fen-Lian, Li, Jin-Song, Zhang, Qian, Zheng, Li-Shu
• Format: Article
• Language: English
• Subjects: Affinity chromatography; Ammonium; Ammonium sulfate; Animals; Antibodies; Antiviral agents; Antiviral Agents - pharmacology; Biological activity; Cell culture; Cell Death - drug effects; Cell Proliferation - drug effects; CHO Cells; Chromatography; Clone Cells; Cloning; Cricetulus; Cytotoxicity; Flp-in-CHO cell; Genetic Vectors - metabolism; Influenza; Interferon; Interferons; Interleukins - isolation & purification; Interleukins - metabolism; Interleukins - pharmacology; Killer Cells, Natural - drug effects; Killer Cells, Natural - metabolism; Natural killer cells; Plasmids; Protein expression; Proteins; Purification; R&D; Recombinant human IFN-λ1(rhIFN-λ1); Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Recombinant Proteins - pharmacology; Research & development; Short Report; Stomatitis; Transfection
• ISSN: 1425-8153; 1689-1392
• DOI: 10.1186/s11658-017-0057-x

Many studies have investigated the characteristics and biological activities of type III interferon (IFN), finding that it has similar features to type I IFN but also unique actions because it is recognized by a different receptor. A full-length recombinant human IFN-λ1 (rhIFN-λ1) cDNA was cloned into the pDF expression vector and stably expressed in Flp-In-CHO cells. After four purification steps (ammonium sulfate precipitation, SP Sepharose chromatography, Blue Sepharose 6 fast flow affinity chromatography and molecular sieve chromatography), the rhIFN-λ1 had a purity of about 90% and was found to have the predicted biological activities. The anti-viral activity of rhIFN-λ1 was determined as 10  IU/mg using the vesicular stomatitis virus (WISH-VSV) assay system. The anti-proliferation activity of rhIFN-λ1 was measured using the MTS method and the growth inhibition ratio was 57% higher than that for recombinant human IFN-α2b (rhIFN-α2b) when the rhIFN-λ1 concentration was 1000 IU/ml. rhIFN-λ1 had lower natural killer cell cytotoxicity than rhIFN-α2b. The Flp-In-CHO system is suitable for stably expressing rhIFN-λ1 that possesses the predicted anti-viral, anti-proliferation and natural killer cell cytotoxicity-promoting activities.