Epitope mapping and targeted quantitation of the cardiac biomarker troponin by SID-MRM mass spectrometry.
Proteomics
; 14(11): 1311-21, 2014 Jun.
Article
em En
| MEDLINE
| ID: mdl-24596168
The absolute quantitation of the targeted protein using MS provides a promising method to evaluate/verify biomarkers used in clinical diagnostics. In this study, a cardiac biomarker, troponin I (TnI), was used as a model protein for method development. The epitope peptide of TnI was characterized by epitope excision followed with LC/MS/MS method and acted as the surrogate peptide for the targeted protein quantitation. The MRM-based MS assay using a stable internal standard that improved the selectivity, specificity, and sensitivity of the protein quantitation. Also, plasma albumin depletion and affinity enrichment of TnI by anti-TnI mAb-coated microparticles reduced the sample complexity, enhanced the dynamic range, and further improved the detecting sensitivity of the targeted protein in the biological matrix. Therefore, quantitation of TnI, a low abundant protein in human plasma, has demonstrated the applicability of the targeted protein quantitation strategy through its epitope peptide determined by epitope mapping method.
Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Mapeamento de Epitopos
/
Troponina I
/
Espectrometria de Massas em Tandem
Tipo de estudo:
Evaluation_studies
Limite:
Humans
Idioma:
En
Revista:
Proteomics
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2014
Tipo de documento:
Article
País de afiliação:
Estados Unidos