Construction of a Novel Liver-Targeting Fusion Interferon by Incorporation of aPlasmodiumRegion I-Plus Peptide
| Autor: | Xiaobao Jin, Fujiang Chu, Yanting Huang, Yan Ma, Juan Shen, Han-fang Mei, Jie Wang, Xuemei Lu, Jiayong Zhu |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
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 |
| Zdroj: | BioMed Research International, Vol 2014 (2014) BioMed Research International |
| ISSN: | 2314-6141 2314-6133 |
| DOI: | 10.1155/2014/261631 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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