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Article

Robust Recombinant Expression of Human Placental Ribonuclease Inhibitor in Insect Cells

TargetEx Ltd., Madách I. u. 31/2., H-2120 Dunakeszi, Hungary
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Academic Editors: Zdenko Levarski and Stanislav Stuchlík
Biomolecules 2022, 12(2), 273; https://doi.org/10.3390/biom12020273
Received: 27 December 2021 / Revised: 2 February 2022 / Accepted: 3 February 2022 / Published: 8 February 2022
(This article belongs to the Special Issue Recombinant Enzymes/Proteins in Biotechnology)
Ribonuclease inhibitors (RIs) are an indispensable biotechnological tool for the detection and manipulation of RNA. Nowadays, due to the outbreak of COVID-19, highly sensitive detection of RNA has become more important than ever. Although the recombinant expression of RNase inhibitors is possible in E. coli, the robust expression is complicated by maintaining the redox potential and solubility by various expression tags. In the present paper we describe the expression of RI in baculovirus-infected High Five cells in large scale utilizing a modified transfer vector combining the beneficial properties of Profinity Exact Tag and pONE system. The recombinant RI is expressed at a high level in a fusion form, which is readily cleaved during on-column chromatography. A subsequent anion exchange chromatography was used as a polishing step to yield 12 mg native RI per liter of culture. RI expressed in insect cells shows higher thermal stability than the commercially available RI products (mainly produced in E. coli) based on temperature-dependent RNase inhibition studies. The endotoxin-free RI variant may also be applied in future therapeutics as a safe additive to increase mRNA stability in mRNA-based vaccines. View Full-Text
Keywords: ribonuclease inhibitor; quantitative PCR; baculovirus-insect expression system; high-level soluble expression ribonuclease inhibitor; quantitative PCR; baculovirus-insect expression system; high-level soluble expression
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MDPI and ACS Style

Flachner, B.; Dobi, K.; Benedek, A.; Cseh, S.; Lőrincz, Z.; Hajdú, I. Robust Recombinant Expression of Human Placental Ribonuclease Inhibitor in Insect Cells. Biomolecules 2022, 12, 273. https://doi.org/10.3390/biom12020273

AMA Style

Flachner B, Dobi K, Benedek A, Cseh S, Lőrincz Z, Hajdú I. Robust Recombinant Expression of Human Placental Ribonuclease Inhibitor in Insect Cells. Biomolecules. 2022; 12(2):273. https://doi.org/10.3390/biom12020273

Chicago/Turabian Style

Flachner, Beáta, Krisztina Dobi, Anett Benedek, Sándor Cseh, Zsolt Lőrincz, and István Hajdú. 2022. "Robust Recombinant Expression of Human Placental Ribonuclease Inhibitor in Insect Cells" Biomolecules 12, no. 2: 273. https://doi.org/10.3390/biom12020273

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