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Construction of a Novel Liver-Targeting Fusion Interferon by Incorporation of aPlasmodiumRegion I-Plus Peptide
- Source :
- BioMed Research International, Vol 2014 (2014), BioMed Research International
- Publication Year :
- 2014
- Publisher :
- Hindawi Limited, 2014.
-
Abstract
- Interferon alpha (IFNα) exerts a multiplicity of biological actions including antiviral, immunomodulatory, and antiproliferative effects. Administration of IFNαis the current treatment for chronic hepatitis B; however, therapy outcome has not been completely satisfactory. The systemic effects of IFNαmay account for its lowin vivobiological activity and multiple adverse events. The purpose of this study was to design a novel liver-targeting fusion interferon (IFN-CSP) by fusing IFNα2b with aPlasmodiumregion I-plus peptide, thus targeting the drug specifically to the liver. The DNA sequence encoding IFN-CSP was constructed using improved splicing by overlapping extension-PCR method, and then cloned into the pET-21b vector for protein expression inE. coliBL21 (DE3). The recombinant protein was expressed as a His-tagged protein and purified using a combination of Ni affinity and HiTrap affinity chromatography at a purity of over 95%. The final yield of biologically active IFN-CSP was up to 270 mg/L culture. The purified recombinant protein showed anti-HBV activity and liver-targeting potentialityin vitro. These data suggests that the novel fusion interferon IFN-CSP may be an excellent candidate as a liver-targeting anti-HBV agent.
- Subjects :
- DNA Replication
Hepatitis B virus
Plasmodium
Article Subject
Recombinant Fusion Proteins
Genetic Vectors
Protozoan Proteins
lcsh:Medicine
Alpha interferon
Peptide
Biology
General Biochemistry, Genetics and Molecular Biology
Cell Line
law.invention
Mice
Affinity chromatography
Interferon
law
In vivo
medicine
Animals
Humans
Chromatography, High Pressure Liquid
chemistry.chemical_classification
Chromatography, Reverse-Phase
Mice, Inbred BALB C
General Immunology and Microbiology
lcsh:R
Interferon-alpha
Biological activity
General Medicine
Molecular biology
In vitro
Liver
chemistry
Organ Specificity
DNA, Viral
Recombinant DNA
Electrophoresis, Polyacrylamide Gel
Peptides
Research Article
Plasmids
medicine.drug
Subjects
Details
- ISSN :
- 23146141 and 23146133
- Volume :
- 2014
- Database :
- OpenAIRE
- Journal :
- BioMed Research International
- Accession number :
- edsair.doi.dedup.....1661d842edd05f1e774ed538b3e7b58c