PO0021 Selective Enrichment of Peptides by Utilizing Dispersive Pipette Extraction on AutomatedLiquid Handler for High Throughput Discovery and Processing
来源:IMCS
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作者:Sunil Hwang, Todd Mullis,L. Andrew Lee*
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发布时间: 1714天前
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1716 次浏览
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According to a database established from human hemofiltrate, approximately 5000 different peptides were recorded. Over 95% of the detected masses were smaller than 15,000 Da and a small percentage (7%) of the detected masses represented sequences frompeptide hormones, growth factors and cytokines. Tese low molecular weight (LMW) peptides/proteins are attractive targets for monitoring human health. Here, we demonstratea proof of concept size exclusion chromatography technique using INtip, dispersive pipetteextraction chemistries (IMCStips™) for rapid isolation of LMW peptides(Bradykinin and Angiotensin I) by using various pore sizes ranging from 30 Å to 60 Å.
INTRODUCTION
Solid phase extraction relies on physicochemical properties in order to partition selectedanalytes from sample solutions. Majority, if not all SPE products are limited in itscapability for purifying and enriching peptides (1-10 kDa) from large pool of interferingproteins (>10 kDa) or small molecules (< 1 kDa). In this study, IMCStips are used toselectively enrich for peptides from biofluids on a 96-channel liquid handler, Microlab®NIMBUS® from Hamilton Company. Mulitvariant factors are imposed in highthroughput manner to optimize the isolation conditions, and thereafter the highthroughput for selected peptides is achieved. Transferring the sample preparation processwith IMCStips required no additional equipment to the automated or semi-automatedliquid handling system.
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Solid phase extraction relies on physicochemical properties in order to partition selectedanalytes from sample solutions. Majority, if not all SPE products are limited in itscapability for purifying and enriching peptides (1-10 kDa) from large pool of interferingproteins (>10 kDa) or small molecules (< 1 kDa). In this study, IMCStips are used toselectively enrich for peptides from biofluids on a 96-channel liquid handler, Microlab®NIMBUS® from Hamilton Company. Mulitvariant factors are imposed in highthroughput manner to optimize the isolation conditions, and thereafter the highthroughput for selected peptides is achieved. Transferring the sample preparation processwith IMCStips required no additional equipment to the automated or semi-automatedliquid handling system.
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