Biologically active, magnICON®-expressed EPO-Fc from stably transformed Nicotiana benthamiana plants presenting tetra-antennary N-glycan structures

J Biotechnol. 2012 Aug 31;160(3-4):242-50. doi: 10.1016/j.jbiotec.2012.03.003. Epub 2012 Mar 16.

Abstract

In the past two decades plants have emerged as a valuable alternative for the production of pharmaceutical proteins. Since N-glycosylation influences functionality and stability of therapeutic proteins, the plant N-glycosylation pathway should be humanized. Here, we report the transient magnICON(®) expression of the erythropoietin fusion protein (EPO-Fc) in Nicotiana benthamiana plants that produce multi-antennary N-glycans without the plant-specific β1,2-xylose and α1,3-fucose residues in a stable manner (Nagels et al., 2011). The EPO-Fc fusion protein consists of EPO with a C-terminal-linked IgG-Fc domain and is used for pulmonary delivery of recombinant EPO to patients (Bitonti et al., 2004). Plant expressed EPO-Fc was quantified using a paramagnetic-particle chemiluminescent immunoassay and shown to be active in vitro via receptor binding experiments in HEK293T cells. Mass spectrometry-based N-glycan analysis confirmed the presence of multi-antennary N-glycans on plant-expressed EPO-Fc. The described research is the next step towards the development of a production platform for pharmaceutical proteins in plants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Erythropoietin / biosynthesis*
  • Erythropoietin / genetics
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / metabolism*
  • Nicotiana / physiology*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / metabolism*
  • Polysaccharides / genetics
  • Polysaccharides / metabolism*
  • Protein Engineering / methods*
  • Transformation, Genetic / genetics

Substances

  • Immunoglobulin Fragments
  • Polysaccharides
  • Erythropoietin