Journal
ACS APPLIED MATERIALS & INTERFACES
Volume 10, Issue 1, Pages 1324-1333Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsami.7b13640
Keywords
PNIPAM; MALDI-MS; photoclick chemistry; surface patterning; on-plate desalting and enrichment
Funding
- National Natural Science Foundation of China [21605083, 21635005]
- Natural Science Foundation of Jiangsu Province [BK20160644]
- National Research Foundation for Thousand Youth Talents Plan of China
- National Science Foundation [CHE-1413449]
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Sample desalting and concentration are crucial steps before matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) analysis. Current sample pretreatment approaches require tedious fabrication and operation procedures, which are unamenable to high-throughput analysis and also result in sample loss. Here, we report the development of a smart MALDI substrate for on-plate desalting, enrichment, and direct MS analysis of protein digests based on thermoresponsive, hydrophilic/hydrophobic transition of surface-grafted poly(N-isopropylacrylamide) (PNIPAM) microarrays. Superhydrophilic 1-thioglycerol microwells are first constructed on alkyne-silane-functionalized rough indium tin oxide substrates based on two sequential thiol-yne photoclick reactions, whereas the surrounding regions are modified with hydrophobic 1H,1H,2H,2H-perfluorodecanethiol. Surface-initiated atom-transfer radical polymerization is then triggered in microwells to form PNIPAM arrays, which facilitate sample loading and enrichment of protein digests by concentrating large-volume samples into small dots and achieving on-plate desalting through PNIPAM configuration change at elevated temperature. The smart MALDI plate shows high performance for mass spectrometric analysis of cytochrome c and neurotensin in the presence of 1 M urea and 100 mM NaHCO3, as well as improved detection sensitivity and high sequence coverage for alpha-casein and cytochrome c digests in femtomole range. The work presents a versatile sample pretreatment platform with great potential for proteomic research.
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