ISSN 2410-7751 (Print)
ISSN 2410-776X (Online)
Biotechnologia Acta Т. 17, No. 2 , 2024
P. 54-55, Bibliography 4, Engl.
UDC::579.6:663.18.
DOI:https://doi.org/10.15407/biotech17.02.054
USE OF SUMO-EXPRESSION SYSTEM AND SUMO-PROTEASE FOR PRODUCTION OF ACTIVE INTERFERON α-2b
I-M.M. Klymkovych 1, 2, M.M. SKRYNNYK 2
1 Educational and Scientific Center “Institute of Biology and Medicine” of Taras Shevchenko National University of Kyiv, Ukraine
2 Department of Research and Development, JSC “Farmak”, Kyiv, Ukraine
Recombinant proteins production in prokaryotic expression systems is often complicated by need of native, N-terminal formylmethionine free molecule extraction, refolding, and processing. To facilitate these stages, alternative expression systems and fusion of target proteins are being used, in particular the SUMO system.
Aim. This research aimed to verify a feasibility of SUMO-protease and SUMO-expression system using to produce active interferon α-2b. The goal was to increase both expression and total yield of the target protein and enhance its quality characteristics, particularly its purity and medical safety, for potential application in acute respiratory diseases treating.
Methods. The obtained SUMO-protease and SUMO-interferon hybrid protein, as well as the proteolytic reaction were analyzed by SDS-PAGE and HPLC techniques.
The results confirmed that SUMO system appliance enhanced protein expression level, refolding efficiency; as well, it was useful for purification costs reduction.
Conclusion. The proteolytic reaction efficiency was close to 100 %.
Key words: SUMO-protease, SUMO-interferon, SUMO-tag, interferon α-2b
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© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2024