title: Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS.
abstract: BACKGROUND
  In this paper we describe a novel method to achieve high yield bacterial expression of a small protein domain with considerable therapeutic potential; Domain I of Beta-2-glycoprotein I (β2GPI). β2GPI is intrinsic to the pathological progression of the Antiphospholipid Syndrome (APS). Patients develop autoantibodies targeting an epitope located on the N-terminal Domain I of β2GPI rendering this domain of interest as a possible therapeutic.
  
  
  RESULTS
  This new method of production of Domain I of β2GPI has increased the production yield by ~20 fold compared to previous methods in E.coli. This largely scalable, partially automated method produces 50-75 mg of pure, folded, active Domain I of β2GPI per litre of expression media.
  
  
  CONCLUSIONS
  The application of this method may enable production of Domain I on sufficient scale to allow its use as a therapeutic.
authors.affiliation: Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, London, WC1E 6JF, UK. Thomas.mcdonnell.11@ucl.ac.uk.
authors.forename: Thomas
authors.initials: T
authors.lastname: McDonnell
authors.affiliation: Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, London, WC1E 6JF, UK. c.pericleous@ucl.ac.uk.
authors.forename: Charis
authors.initials: C
authors.lastname: Pericleous
authors.affiliation: PolyTherics, Babraham Research Campus, Babraham, CB22 3AT, Cambridge, UK. elaurine@hotmail.com.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Laurine
authors.affiliation: PolyTherics, Babraham Research Campus, Babraham, CB22 3AT, Cambridge, UK. Rita.Tommasi@gmail.com.
authors.forename: Rita
authors.initials: R
authors.lastname: Tommasi
authors.affiliation: Structural Biology, Medical Research Council National Institute for Medical Research, London, UK. acely.garza@crick.ac.uk.
authors.forename: Acely
authors.initials: A
authors.lastname: Garza-Garcia
authors.affiliation: Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, London, WC1E 6JF, UK. i.giles@ucl.ac.uk.
authors.forename: Ian
authors.initials: I
authors.lastname: Giles
authors.affiliation: Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, London, WC1E 6JF, UK. y.ioannou@ucl.ac.uk.|Arthritis Research UK Centre for Adolescent Rheumatology, University College London, London, UK. y.ioannou@ucl.ac.uk.
authors.forename: Yiannis
authors.initials: Y
authors.lastname: Ioannou
authors.affiliation: Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, London, WC1E 6JF, UK. Anisur.rahman@ucl.ac.uk.
authors.forename: Anisur
authors.initials: A
authors.lastname: Rahman
mesh_terms: D000328:Adult; D016736:Antiphospholipid Syndrome / Q000188:drug therapy*; D057205:Automation, Laboratory; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D006801:Humans; D002479:Inclusion Bodies; D008297:Male; D008875:Middle Aged; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000627:therapeutic use; D053482:beta 2-Glycoprotein I / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism* / Q000627:therapeutic use
chemical_list: D011993:Recombinant Fusion Proteins; D053482:beta 2-Glycoprotein I