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Proteomics ; 14(21-22): 2432-6, 2014 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-25263329

RESUMEN

DNA-assisted proteomics technologies enable ultra-sensitive measurements in multiplex format using DNA-barcoded affinity reagents. Although numerous antibodies are available, nowadays targeting nearly the complete human proteome, the majority is not accessible at the quantity, concentration, or purity recommended for most bio-conjugation protocols. Here, we introduce a magnetic bead-assisted DNA-barcoding approach, applicable for several antibodies in parallel, as well as reducing required reagents quantities up to a thousand-fold. The success of DNA-barcoding and retained functionality of antibodies were demonstrated in sandwich immunoassays and standard quantitative Immuno-PCR assays. Specific DNA-barcoding of antibodies for multiplex applications was presented on suspension bead arrays with read-out on a massively parallel sequencing platform in a procedure denoted Immuno-Sequencing. Conclusively, human plasma samples were analyzed to indicate the functionality of barcoded antibodies in intended proteomics applications.


Asunto(s)
Anticuerpos/química , Proteínas Sanguíneas/análisis , ADN/química , Proteómica/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Humanos , Inmunoensayo/métodos , Inmunoconjugados/química , Separación Inmunomagnética/métodos , Reacción en Cadena de la Polimerasa/métodos
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