Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS

BMC Biotechnology

Methodology Article

Charis Pericleous¹  Anisur Rahman¹  Ian Giles¹  Thomas McDonnell¹  Yiannis Ioannou²  Emmanuelle Laurine³  Rita Tommasi³  Acely Garza-Garcia⁴ 

[1] Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, WC1E 6JF, London, UK;Centre for Rheumatology, Division of Medicine, University College London, Rayne Institute, 5 University Street, WC1E 6JF, London, UK;Arthritis Research UK Centre for Adolescent Rheumatology, University College London, London, UK;PolyTherics, Babraham Research Campus, Babraham, CB22 3AT, Cambridge, UK;Structural Biology, Medical Research Council National Institute for Medical Research, London, UK;

关键词: Antiphospholipid syndrome;    Protein production;    Inclusion bodies;    E. Coli;    Beta-2-Glycoprotein I;    Domain I;    Automated;   

DOI  :  10.1186/s12896-015-0222-0

 received in 2015-04-28, accepted in 2015-11-03,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundIn 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.ResultsThis 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.ConclusionThe application of this method may enable production of Domain I on sufficient scale to allow its use as a therapeutic.

【 授权许可】

CC BY   
© McDonnell et al. 2015