title: Construction and characterization of a potent, long-lasting recombinant human serum albumin-interferon α1 fusion protein expressed in Pichia pastoris.
abstract: The main obstacles to using Interferon a1 as an antiviral agent are its low stability and fast clearance. Here, we prepared a long-lasting recombinant human serum albumin-interferon α1 fusion protein. It was expressed in methylotrophic yeast Pichia pastoris with HSA's natural signal peptide and purified by dye affinity chromatography and ion exchange chromatography. The physicochemical, biological, and pharmaceutical characteristics of HSA-IFN α1 were then evaluated in vitro and in vivo. The purity of HSA-IFN α1 was about 95% analyzed by SDS-PAGE. Molecular weight determined by MALDI-TOF mass spectrometry was 86,582. Western blot analysis showed that the expressed HSA-IFN α1 had the antigenicity of HSA. The N-terminal acid amino sequence was identical to predicted sequence. The antivirus activity in vitro evaluated by cytopathic effect assay was (1.63±0.06)×10(5) IU/mg. After administered in rats, the biological activity of HSA-IFN α1 were enhanced and maintained for 6days in serum. Overall, these studies demonstrate the feasibility of using albumin-fusion technology to development a long-lasting, antiviral protein HSA-IFN α1 with high antivirus activity.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100005, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Tian
authors.forename: Qinshan
authors.initials: Q
authors.lastname: Li
authors.forename: Wenbin
authors.initials: W
authors.lastname: Yao
authors.forename: Chen
authors.initials: C
authors.lastname: Xu
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics*; D008970:Molecular Weight; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D012709:Serum Albumin / Q000235:genetics*
chemical_list: D000998:Antiviral Agents; C554156:IFNA1 protein, human; D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin
keywords: Antiviral activity; Interferon; Long-lasting; Pichia pastoris; Purification