High throughput peptide mass fingerprinting and protein macroarray analysis using chemical printing strategies.
Mol Cell Proteomics
; 1(7): 490-9, 2002 Jul.
Article
em En
| MEDLINE
| ID: mdl-12239277
ABSTRACT
We describe a chemical printer that uses piezoelectric pulsing for rapid, accurate, and non-contact microdispensing of fluid for proteomic analysis of immobilized protein macroarrays. We demonstrate protein digestion and peptide mass fingerprinting analysis of human plasma and platelet proteins direct from a membrane surface subsequent to defined microdispensing of trypsin and matrix solutions, hence bypassing multiple liquid-handling steps. Detection of low abundance, alkaline proteins from whole human platelet extracts has been highlighted. Membrane immobilization of protein permits archiving of samples pre-/post-analysis and provides a means for subanalysis using multiple chemistries. This study highlights the ability to increase sequence coverage for protein identification using multiple enzymes and to characterize N-glycosylation modifications using a combination of PNGase F and trypsin. We also demonstrate microdispensing of multiple serum samples in a quantitative microenzyme-linked immunosorbent assay format to rapidly screen protein macroarrays for pathogen-derived antigens. We anticipate the chemical printer will be a major component of proteomic platforms for high throughput protein identification and characterization with widespread applications in biomedical and diagnostic discovery.
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Base de dados:
MEDLINE
Assunto principal:
Mapeamento de Peptídeos
/
Proteoma
/
Proteômica
Limite:
Humans
Idioma:
En
Ano de publicação:
2002
Tipo de documento:
Article