title: 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.
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.
authors.affiliation: Department of Pharmacy, University of Patras, Patras, Greece.
authors.forename: Athanasios
authors.initials: A
authors.lastname: Niarchos
authors.affiliation: Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
authors.forename: Marios
authors.initials: M
authors.lastname: Zouridakis
authors.affiliation: Institute of Biosciences and Applications, National Centre for Scientific Research "Demokritos", Athens, Greece.
authors.forename: Vassilis
authors.initials: V
authors.lastname: Douris
authors.affiliation: Department of Pharmacy, University of Patras, Patras, Greece.
authors.forename: Assimina
authors.initials: A
authors.lastname: Georgostathi
authors.affiliation: Department of Pharmacy, University of Patras, Patras, Greece.
authors.forename: Dimitra
authors.initials: D
authors.lastname: Kalamida
authors.affiliation: Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
authors.forename: Alexandros
authors.initials: A
authors.lastname: Sotiriadis
authors.affiliation: Department of Pharmacy, University of Patras, Patras, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Poulas
authors.affiliation: Institute of Biosciences and Applications, National Centre for Scientific Research "Demokritos", Athens, Greece.
authors.forename: Kostas
authors.initials: K
authors.lastname: Iatrou
authors.affiliation: Department of Pharmacy, University of Patras, Patras, Greece ; Department of Biochemistry, Hellenic Pasteur Institute, Athens, Greece.
authors.forename: Socrates J
authors.initials: SJ
authors.lastname: Tzartos
mesh_terms: D000818:Animals; D001323:Autoantibodies / Q000097:blood / Q000378:metabolism*; D015153:Blotting, Western; D002038:Bungarotoxins / Q000378:metabolism; D002460:Cell Line; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D007163:Immunosorbent Techniques; D007457:Iodine Radioisotopes / Q000378:metabolism; D009036:Moths; D018482:Muscle, Skeletal / Q000378:metabolism*; D009157:Myasthenia Gravis / Q000276:immunology* / Q000628:therapy*; D010843:Pichia; D010957:Plasmids / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000502:physiology; D011863:Radioimmunoassay; D011978:Receptors, Nicotinic / Q000378:metabolism* / Q000627:therapeutic use*
chemical_list: D001323:Autoantibodies; D002038:Bungarotoxins; C523021:CHRNA1 protein, human; D000939:Epitopes; D007457:Iodine Radioisotopes; D011978:Receptors, Nicotinic