Production of Recombinant IFN-α2b from Tobacco (Nicotiana tobacum) by Non-Chromatography Purification

Abstract

Developing genetically modified plants has been among wildly used strategies to produce recombinant proteins. Prior to have a large-scale production of a recombinant proteins, transient expression in a model plant is recommended as it could lead us to a proper insight of the subject. In this research, interferon alpha 2b gene (INF α2b) as a protein with antiviral and antitumor activities was subjected to be transiently expressed in tobacco and through Agrobacterium tumefaciens infiltration method. The optimized target sequence, IFN α2b, was designed to be constructed in such a way that it could be purified either by His-tag or Elastin-like peptide (ELP). Agro-infiltrated leaves were analyzed through real time PCR. In proteomics level, the ELP-tagged IFN was successfully purified through non-chromatography method and confirmed by SDS-PAGE electrophoresis. Considering the low level of recombinant protein production in the plants, one remarkable achievement of this research was application of ELP in combination with IFN α2b for purification of recombinant protein from total protein of the plant hosts.