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

开发用于治疗APS的β-2-糖蛋白I结构域I的高产量表达和纯化系统

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作者:Thomas McDonnell, Charis Pericleous, Emmanuelle Laurine, Rita Tommasi, Acely Garza-Garcia, Ian Giles, Yiannis Ioannou, Anisur Rahman

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.

Conclusion

The application of this method may enable production of Domain I on sufficient scale to allow its use as a 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.

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