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| Expression of Human Serum Albumin-interferon α2b Fusion Protein in Chinese Hamster Ovary Cells |
| LI Cheng-yuan1, ZHANG Jing-jing1, QIAN Kai2, MIAO Ya-na1, CAI Yan-fei1, YANG Jian-feng3, HE Yang4, JIN Jian1 |
1 School of Pharmaceutical Sciences, Jiangnan University, Wuxi 214122, China;
2 College of Life and Environmental Science, Wenzhou University, Wenzhou 325035, China;
3 Cyrus Tang Hematology Center and Ministry of Education Engineering Center of Hematological Disease, Soochow University, Suzhou 215123, China;
4 Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou 215006, China |
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Abstract Interferon α2b (IFNα2b) is a multifunctional cytokine for the treatment of viral diseases and tumor diseases, and its clinical applications are limited by the short half-life. IFNα2b was fused to the C-terminus of human serum albumin (HSA) and the pMH3 plasmid containing fusion protein (HSA-IFNα2b) encoding gene was constructed. Fusion protein expression Chinese hamster ovary (CHO) cell line was established by electroporation and high expression cell lines were screened by adding G418. The expressed fusion protein was analyzed by Western blot and results showed that the expressed protein was detected by both anti-bodies of HSA and IFNα2b. In suspension culture, fusion protein concentration achieved 65mg/L at the end of batch fermentation. Different generations of fusion expression cell lines showed the similar cell growth and protein production and thus a stable expression cell line was obtained. In 3-L fed-batch culture, the optimized culture time was 15 days and final HSA-IFNα2b production was 121mg/L. After two-step purification, the purity of fusion protein was 96.8% and the total yield was 22.3%. According to the "Chinese Pharmacopoeia 2015 edition" of the IFNα2b detection method, the results show that the anti-virus activity is 4.1x106IU/mg. In conclusion, fusion protein of HSA-IFNα2b was first expressed in mammalian cell CHO and the obtained fusion had high biological activity.
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Received: 13 January 2016
Published: 25 July 2016
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