Functional expression of a biologically active fragment of soluble gp130 as an ELP-fusion protein in transgenic plants: purification via inverse transition cycling

Meng Lin , Stefan Rose-John , Joachim Grötzinger , Udo Conrad , Jürgen Scheller , Jürgen Scheller (1)

1 Biochemisches Institut

In murine models of Crohn‘s disease, rheumatoid arthritis and colon cancer, interleukin (IL)-6 signalling via the soluble IL-6 receptor (sIL-6R, termed IL-6 trans-signalling) has been shown to promote the pathology associated with these conditions. These detrimental activities can however be selectively blocked by soluble forms of the gp130 receptor. Although soluble gp130 (sgp130) therefore represents a viable therapeutic modality for the treatment of these conditions, the mass manufacture of such biologics is often expensive. The advent of molecular farming has however provided an extremely cost effective strategy for the engineering of recombinant proteins. Here we describe the expression and production of a biologically active sgp130 variant which is expressed in transgenic tobacco plants as an elastin-like pentapeptide (ELP) fusion protein (mini-gp130-ELP). Mini-gp130-ELP consists of the first three domains of gp130 (Ig-like domain and cytokine binding module) fused to 100 repeats of ELP. Expression of mini-gp130-ELP did not affect the growth rate or morphology of the transgenic plants, and purification was achieved using inverse transition cycling. This approach led to an overall yield of 141 µg of purified protein per gram of fresh leaf weight. The purified mini-gp130-ELP specifically inhibited sIL-6R-mediated trans-signalling as measured by binding to the IL-6/sIL-6R complex and through its ability to block sIL-6R-mediated activation of STAT3 phosphorylation and proliferation in human hepatoma cells and murine pre-B-cells. Consequently, this study validates the potential application of molecular farming in transgenic tobacco plants as a strategy for the expression and purification of therapeutically advantageous biologics such as sgp130.

Mots clés: Life Sciences

Meng Lin, Stefan Rose-John, Joachim Grötzinger, Udo Conrad, Jürgen Scheller, et al.. Functional expression of a biologically active fragment of soluble gp130 as an ELP-fusion protein in transgenic plants: purification via inverse transition cycling. Biochemical Journal, 2006, 398 (3), pp.577-583. ⟨10.1042/BJ20060544⟩. ⟨hal-00478575⟩