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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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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
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author McDonnell, Thomas
Pericleous, Charis
Laurine, Emmanuelle
Tommasi, Rita
Garza-Garcia, Acely
Giles, Ian
Ioannou, Yiannis
Rahman, Anisur
author_facet McDonnell, Thomas
Pericleous, Charis
Laurine, Emmanuelle
Tommasi, Rita
Garza-Garcia, Acely
Giles, Ian
Ioannou, Yiannis
Rahman, Anisur
author_sort McDonnell, Thomas
collection PubMed
description 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.
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spelling pubmed-46502792015-11-19 Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS McDonnell, Thomas Pericleous, Charis Laurine, Emmanuelle Tommasi, Rita Garza-Garcia, Acely Giles, Ian Ioannou, Yiannis Rahman, Anisur BMC Biotechnol Methodology Article 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. BioMed Central 2015-11-14 /pmc/articles/PMC4650279/ /pubmed/26576675 http://dx.doi.org/10.1186/s12896-015-0222-0 Text en © McDonnell et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
McDonnell, Thomas
Pericleous, Charis
Laurine, Emmanuelle
Tommasi, Rita
Garza-Garcia, Acely
Giles, Ian
Ioannou, Yiannis
Rahman, Anisur
Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS
title Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS
title_full Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS
title_fullStr Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS
title_full_unstemmed Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS
title_short Development of a high yield expression and purification system for Domain I of Beta-2-glycoprotein I for the treatment of APS
title_sort development of a high yield expression and purification system for domain i of beta-2-glycoprotein i for the treatment of aps
topic Methodology Article
url 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
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