Expression of a highly antigenic and native-like folded extracellular domain of the human α1 subunit of muscle nicotinic acetylcholine receptor, suitable for use in antigen specific therapies for Myasthenia Gravis

PLoS One. 2013 Dec 20;8(12):e84791. doi: 10.1371/journal.pone.0084791. eCollection 2013.

Abstract

We describe the expression of the extracellular domain of the human α1 nicotinic acetylcholine receptor (nAChR) in lepidopteran insect cells (i-α1-ECD) and its suitability for use in antigen-specific therapies for Myasthenia Gravis (MG). Compared to the previously expressed protein in P. pastoris (y-α1-ECD), i-α1-ECD had a 2-fold increased expression yield, bound anti-nAChR monoclonal antibodies and autoantibodies from MG patients two to several-fold more efficiently and resulted in a secondary structure closer to that of the crystal structure of mouse α1-ECD. Our results indicate that i-α1-ECD is an improved protein for use in antigen-specific MG therapeutic strategies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoantibodies / blood
  • Autoantibodies / metabolism*
  • Blotting, Western
  • Bungarotoxins / metabolism
  • Cell Line
  • Circular Dichroism
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes
  • Humans
  • Immunosorbent Techniques
  • Iodine Radioisotopes / metabolism
  • Moths
  • Muscle, Skeletal / metabolism*
  • Myasthenia Gravis / immunology*
  • Myasthenia Gravis / therapy*
  • Pichia
  • Plasmids / genetics
  • Protein Structure, Tertiary / physiology
  • Radioimmunoassay
  • Receptors, Nicotinic / metabolism*
  • Receptors, Nicotinic / therapeutic use*

Substances

  • Autoantibodies
  • Bungarotoxins
  • CHRNA1 protein, human
  • Epitopes
  • Iodine Radioisotopes
  • Receptors, Nicotinic

Grants and funding

This work was supported by grants from the European Commission (FP7 FightMG no. 242210, and FP7 REGPOT 2010-1 NeuroSign no. 264083) and the Greek NSRF “Thalis” Autoimmunity grant. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.