High-Throughput Bioassays using "Dip-and-Go" Multiplexed Electrospray Mass Spectrometry.
Angew Chem Int Ed Engl
; 58(49): 17594-17598, 2019 12 02.
Article
em En
| MEDLINE
| ID: mdl-31589796
ABSTRACT
A multiplexed system based on inductive nanoelectrospray mass spectrometry (nESI-MS) has been developed for high-throughput screening (HTS) bioassays. This system combines inductive nESI and field amplification micro-electrophoresis to achieve a "dip-and-go" sample loading and purification strategy that enables nESI-MS based HTS assays in 96-well microtiter plates. The combination of inductive nESI and micro-electrophoresis makes it possible to perform efficient in situ separations and clean-up of biological samples. The sensitivity of the system is such that quantitative analysis of peptides from 1-10 000â
nm can be performed in a biological matrix. A prototype of the automation system has been developed to handle 12 samples (one row of a microtiter plate) at a time. The sample loading and electrophoretic clean-up of biosamples can be done in parallel within 20â
s followed by MS analysis at a rate of 1.3 to 3.5â
s per sample. The system was used successfully for the quantitative analysis of BACE1-catalyzed peptide hydrolysis, a prototypical HTS assay of relevance to drug discovery. IC50 values for this system were in agreement with LC-MS but recorded in times more than an order of magnitude shorter.
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Base de dados:
MEDLINE
Assunto principal:
Peptídeos
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Secretases da Proteína Precursora do Amiloide
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Ensaios de Triagem em Larga Escala
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article