Clean and Folded: Optimized Production of High Quality Recombinant Human Interferon-Λ1

Abstract

Type III interferons exhibit antiviral activity against influenza viruses, coronaviruses, rotaviruses, and others. In addition, this type of interferon theoretically has therapeutic advantages, in comparison with type I interferons, due to its ability to activate a narrower group of genes in a relatively small group of target cells. Hence, it can elicit more targeted antiviral or immunomodulatory responses. Obtaining biologically-active interferon lambda (hIFN-λ1) is fraught with difficulties at the stage of expression in soluble form or, in the case of expression in the form of inclusion bodies, at the stage of refolding. In this work, hIFN-λ1 was expressed in the form of inclusion bodies, and a simple, effective refolding method was developed. Efficient and scalable methods for chromatographic purification of recombinant hIFN-λ1 were also developed. High-yield, high-purity product was obtained through optimization of several processes including: recombinant protein expression; metal affinity chromatography; cation exchange chromatography; and an intermediate protein refolding stage. The obtained protein was shown to have expected, specific biological activity in line with published effects: induction of MxA gene expression in A549 cells.

IFN-λ1 Displays Various Levels of Antiviral Activity In Vitro in a Select Panel of RNA Viruses

“…The presented study focuses on features that relate to the use of human IFN-λ1 as a potential antiviral agent against specific respiratory viruses. We have previously obtained recombinant human interferon-λ1 [26]. It is known that both IFN-α/β and IFNλ induce the expression of similar groups of ISGs (interferon stimulated genes) [21,28], which include such canonical antiviral genes as MxA, OAS, PKR, IFITM, and ISG15 [29,30].…”

“…To study the potential antiviral effects of type III interferon, we used recombinant (human) IFN-lambda protein previously obtained and characterized by the authors [26]. The amino acid sequence of recombinant 6× His-hIFN-λ1 protein was confirmed using MALDI-TOF mass spectrometry.…”