Expression of human interferon alpha-1 with enhanced stability via the tagging system of a stabilizing peptide |
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Authors: | Kim Young Mok Lee Hye Ja Lee Jung Eun Kim Hae Yeong Kim Jongsun |
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Institution: | aATGen Inc., Sung Nam, Kyung Ki Do, South Korea;bDepartment of Food Science and Biotechnology, KyungHee University, Yongin 449-701, Kyung Ki Do, South Korea;cDepartment of Microbiology and Brain Korea 21 Project for Medical Sciences, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, 134 Shinchon-dong, Seodaemoon-gu, Seoul 120-175, Republic of Korea |
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Abstract: | Human interferon α-1 (hIFNA1) is one of several interferon α subtypes that have been studied and commercialized to treat various viral diseases including hepatitis B and C as well as malignant melanoma. Protein aggregation has been problematic for every step in commercial production, from purification to the packaging and delivery of pharmaceutical proteins. In a previous study, we demonstrated that a stabilizing peptide from the C-terminal acidic tail of α-synuclein (ATS) could be used as an effective fusion tag to increase the stability of target proteins such as human growth hormone (hGH) and granulocyte colony-stimulating factor (G-CSF). In this study, we applied this ATS fusion system to hIFNA1 in order to protect against the aggregation of hIFNA1 by environmental stresses, since hIFNA1 aggregates elicit an undesirable immune response in humans. As expected, ATS-fused hIFNA1 (hIFNA1-ATS) protein showed enhanced stability against thermal stress, agitation stress, and repetitive freeze/thawing stress in comparison with native hIFNA1. More importantly, hIFNA1-ATS fusion protein appeared to be 1.6 times more active than hIFNA1 in a cell anti-proliferation assay. Furthermore, the solubility of hIFNA1-ATS appeared to be 1.7 times higher than that of native protein. Our results suggest that the ATS-tag system could be a useful means for protecting hIFNA1 protein from aggregation by various external stresses and could be used to increase the solubility of protein. |
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Keywords: | Interferon alpha Protein aggregation Environmental stresses Stability Protein solubility |
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