Proteomics

Dataset Information

35

Mycobacterium tuberculosis protein production pipeline


ABSTRACT: Purpose: Tuberculosis is still a major threat to global health. New tools and strategies to produce disease-related proteins are quintessential for the development of novel vaccines and diagnostic markers. Experimental design: To obtain recombinant proteins from Mycobacterium tuberculosis (Mtb) for use in clinical applications, a standardized procedure was developed that includes subcloning, protein expression in Mycobacterium smegmatis and protein purification using chromatography. The potential for the different protein targets to serve as diagnostic markers for tuberculosis was established using multiplex immunoassays. Results: Twelve soluble proteins from Mtb, including one protein complex, were purified to homogeneity following recombinant expression in M. smegmatis. Protein purity was assessed both by size exclusion chromatography and mass spectrometry. Multiplex serological testing of the final protein preparations showed that all but one protein displayed a clear antibody response in serum samples from 278 tuberculosis patients. Conclusion and clinical relevance: The established workflow comprises a simple, cost-effective and scalable pipeline for production of soluble proteins from Mtb and can be used to prioritize immunogenic proteins suitable for use as diagnostic markers.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mycobacterium tuberculosis H37Rv  

TISSUE(S): Tissue Not Applicable To Dataset

DISEASE(S): Tuberculosis

SUBMITTER: Joanna Kirkpatrick  

LAB HEAD: Annabel Parret

PROVIDER: PXD004133 | Pride | 2018-06-27

REPOSITORIES: Pride

altmetric image

Publications

A standardized production pipeline for high profile targets from Mycobacterium tuberculosis.

Milewski Morlin C MC   Broger Tobias T   Kirkpatrick Joanna J   Filomena Angela A   Komadina Dana D   Schneiderhan-Marra Nicole N   Wilmanns Matthias M   Parret Annabel H A AH  

Proteomics. Clinical applications 20160803 9-10


Tuberculosis is still a major threat to global health. New tools and strategies to produce disease-related proteins are quintessential for the development of novel vaccines and diagnostic markers.To obtain recombinant proteins from Mycobacterium tuberculosis (Mtb) for use in clinical applications, a standardized procedure was developed that includes subcloning, protein expression in Mycobacterium smegmatis and protein purification using chromatography. The potential for the different protein tar  ...[more]

Similar Datasets

2018-10-26 | PXD005765 | Pride
2013-12-03 | E-GEOD-50050 | ArrayExpress
2019-10-04 | E-MTAB-7679 | ExpressionAtlas
2018-08-06 | PXD010659 | Pride
2020-03-12 | E-MTAB-7679 | ArrayExpress
2016-07-06 | PXD002542 | Pride
2015-11-11 | E-GEOD-71200 | ArrayExpress
2015-03-31 | E-GEOD-67427 | ArrayExpress
2014-10-08 | E-GEOD-11199 | ExpressionAtlas
2014-10-01 | E-GEOD-60376 | ArrayExpress