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
The need for native, N-terminal formylmethionine-free molecule extraction, refolding, and processing often complicates recombinant protein production in prokaryotic expression systems. Alternative expression systems and fusion of target proteins are being used to facilitate these stages, particularly the SUMO system.
Aim. This research aimed to verify the feasibility of the SUMO-protease and SUMO-expression systems used to produce active interferon α-2b. The goal was to increase the target protein's expression and total yield and enhance its quality characteristics, particularly its purity and medical safety, for potential application in acute respiratory diseases.
Methods. The SUMO-protease and SUMO-interferon hybrid protein, as well as the proteolytic reaction were analyzed using SDS-PAGE and HPLC techniques.
The results confirmed that the SUMO system appliance enhanced protein expression level and refolding efficiency; it was also helpful in reducing purification costs.
Conclusion. The proteolytic reaction efficiency was close to 100 %.
Key words: SUMO-protease, SUMO-interferon, SUMO-tag, interferon α-2b
© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2024