title: Increased antiviral activity of microscale-purified HuIFN alpha 8 (human interferon alpha 8) over HuIFN alpha 2b in Hep-2 cells challenged with Mengo virus.
abstract: Human proteins are not routinely expressed at high levels in Escherichia coli for, among other reasons, different codon usage. Several purification procedures have been applied to recover recombinant proteins for further biological characterization. However, the vast majority involve costly chromatography procedures. In the present study, both (Hu)IFN(alpha 2b) (human interferon alpha 2b) and (Hu)IFN(alpha 8) were expressed efficiently in E. coli BL21-codonplus-RIL. Subsequently, both recombinant proteins were purified to homogeneity by passive elution from reverse-stained SDS/PAGE gels, a cost-effective purification procedure. After purification, both recovered proteins were biologically active. The (Hu)IFN(alpha 8) subtype induced 1.46-fold more antiviral activity than (Hu)IFN(alpha 2b) using Hep-2 human laryngeal carcinoma cell challenged with Mengo virus.
authors.affiliation: Process Control Department, Center for Genetic Engineering and Biotechnology, Ave. 31 e/ 158 y 190, P.O. Box 6162, C.P. 10600, City of Havana, Cuba. julio.sanchez@cigb.edu.cu
authors.forename: Julio César Sánchez
authors.initials: JC
authors.lastname: García
authors.forename: Alejandro Miranda
authors.initials: AM
authors.lastname: Ariza
authors.forename: Alexis Musacchio
authors.initials: AM
authors.lastname: Lasa
authors.forename: Luis Javier
authors.initials: LJ
authors.lastname: González
authors.forename: Vladimir Besada
authors.initials: VB
authors.lastname: Perez
mesh_terms: D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D045744:Cell Line, Tumor; D004926:Escherichia coli; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D007372:Interferons / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D008574:Mengovirus / Q000187:drug effects* / Q000276:immunology*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins
chemical_list: D000998:Antiviral Agents; C554164:IFNA8 protein, human; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D007372:Interferons