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Soluble expression of recombinant human interleukin-2 in Escherichia coli and its facile production.
- Source :
-
Protein Expression & Purification . Sep2024, Vol. 221, pN.PAG-N.PAG. 1p. - Publication Year :
- 2024
-
Abstract
- Recombinant human interleukin-2 (rhIL-2) represents one of the most difficult-to-produce cytokines in E. coli due to its extreme hydrophobicity and high tendency to formation of inclusion bodies. Refolding of rhIL-2 inclusion bodies always represents cumbersome downstream processes and low production efficiency. Herein, we disclosed a fusion strategy for efficiently soluble expression and facile production of rhIL-2 in E. coli Origami B (DE3) host. A two-tandem SUMO fusion partner (His-2SUMO) with a unique SUMO protease cleavage site at C-terminus was devised to fuse with the N-terminus of rhIL-2 and the fusion protein (His-2SUMO-rhIL-2) was almost completely expressed in a soluble from. The fusion partner could be efficiently removed by Ulp1 cleavage and the rhIL-2 was simply produced by a two-step Ni-NTA affinity chromatography with a considerable purity and whole recovery. The eventually obtained rhIL-2 was well-characterized and the results showed that the purified rhIL-2 exhibits a compact and ordered structure. Although the finally obtained rhIL-2 exists in a soluble aggregates form and the aggregation probably has been occurred during expression stage, the soluble rhIL-2 aggregates remain exhibit comparable bioactivity with the commercially available rhIL-2 drug formulation. • rhIL-2 is a difficult-to-express cytokine because it highly inclines to forming IBs in E. coli. • Soluble expression of rhIL-2 was achieved in E. coli by fusion with a two-tandem SUMO fusion partner. • A two-step Ni-NTA affinity chromatography was established to efficiently purify the rhIL-2. • The resulted rhIL-2 existed as biologically active soluble aggregates with ordered structure. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 221
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 177847518
- Full Text :
- https://doi.org/10.1016/j.pep.2024.106507