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

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). Pat...

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Detalles Bibliográficos
Autores principales: McDonnell, Thomas, Pericleous, Charis, Laurine, Emmanuelle, Tommasi, Rita, Garza-Garcia, Acely, Giles, Ian, Ioannou, Yiannis, Rahman, Anisur
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4650279/
https://www.ncbi.nlm.nih.gov/pubmed/26576675
http://dx.doi.org/10.1186/s12896-015-0222-0
Descripción
Sumario: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. CONCLUSION: The application of this method may enable production of Domain I on sufficient scale to allow its use as a therapeutic.