Tick-borne encephalitis nonstructural protein NS1 expressed in E. coli retains immunological properties of the native protein

There is evidence that flaviviral NS1 glycoprotein plays an important role in the pathology of tick-borne encephalitis (TBE) and NS1-specific antibodies are detected in the blood of patients with TBE. This makes NS1 a good target for the development of therapeutic inhibitors and NS1 could be an impo...

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Bibliographic Details
Published in:Protein expression and purification 2022-03, Vol.191, p.106031-106031, Article 106031
Main Authors: Andrey, Matveev, Yana, Khlusevich, Olga, Golota, Bogdana, Kravchuk, Sergey, Tkachev, Lyudmila, Emelyanova, Nina, Tikunova
Format: Article
Language:English
Subjects:
Antibodies, Viral - chemistry
Antibodies, Viral - immunology
E. coli expression
Encephalitis Viruses, Tick-Borne - chemistry
Encephalitis Viruses, Tick-Borne - genetics
Encephalitis Viruses, Tick-Borne - immunology
Encephalitis Viruses, Tick-Borne - metabolism
Enzyme-Linked Immunosorbent Assay
Escherichia coli - genetics
Escherichia coli - metabolism
Gene Expression
NS1 protein
Recombinant protein
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - immunology
Tick-borne encephalitis
Trx-fused protein
Viral Nonstructural Proteins - biosynthesis
Viral Nonstructural Proteins - chemistry
Viral Nonstructural Proteins - genetics
Viral Nonstructural Proteins - immunology
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Summary:There is evidence that flaviviral NS1 glycoprotein plays an important role in the pathology of tick-borne encephalitis (TBE) and NS1-specific antibodies are detected in the blood of patients with TBE. This makes NS1 a good target for the development of therapeutic inhibitors and NS1 could be an important biomarker for the early diagnosis of TBE in vaccinated individuals. Eukaryotic expression systems are mainly used to produce recombinant tick-borne encephalitis virus (TBEV) NS1. The expression of TBEV NS1 proteins in eukaryotic cells was successful, but there were some limitations. Several attempts have also been made to obtain the NS1 protein in Escherichia coli cells; however, they were unsuccessful due to the low solubility of the recombinant protein and improper folding. In this study, using Trx-tag as a fusion partner, soluble Trx-fused TBEV NS1 protein was first produced in the E. coli BL21 strain. In addition, insoluble Trx-fused TBEV NS1 protein was obtained when cultivation conditions were changed to increase the productivity. The insoluble TBEV NS1 obtained from inclusion bodies was solubilized using chaotropic reagents and successfully refolded using dialysis. Both soluble variant and successfully refolded from inclusion bodies variant showed immunological properties similar to the native TBEV NS1 protein and were recognized by specific monoclonal antibodies (mAbs), immune ascetic fluid in ELISA, western blot, and competitive analysis. •Soluble TBEV NS1 proteins was first expressed in E. coli.•NS1 proteins were expressed using a Trx-tag fusion.•The purified NS1 proteins were antigenically active and recognized by antibodies.•Obtained protein is valuable for the development of diagnostics.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2021.106031