Capillary‐channeled polymer (C‐CP) fibers for the rapid extraction of proteins from urine matrices prior to detection with MALDI‐MS. Issue 5 (18th March 2015)

• Record Type: Journal Article
• Authors: Manard, Benjamin T.; Jones, Sarah M. H.; Marcus, R. Kenneth; Capelo Martínez, José Luis; Lodeiro, Carlos; Santos, Hugo M.
• Is Part Of: Proteomics. Volume 9:Issue 5/6(2015)
• Page Start: 522
• Page End: 530
• Publication Date: 2015-03-18
• Subjects: Proteomics -- Periodicals; 572.605
• DOI: 10.1002/prca.201400081
• Languages: English
• ISSNs: 1862-8346

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="prca1605-sec-0010" sec-type="section"> <title>Purpose</title> <p>While MS is a powerful tool for biomarker determinations, the high salt content and the small molecules present in urine poses incredible challenges. Separation/extraction methods must be employed for the isolation of target species at relevant concentrations. Micropipette tips packed with capillary‐channeled polymer (C‐CP) fibers are employed for the SPE of proteins from a synthetic and a certified urine matrix.</p> </sec> <sec id="prca1605-sec-0020" sec-type="section"> <title>Experimental design</title> <p>Extractions are performed utilizing a very simple centrifugation method to spin‐down species through the C‐CP fiber tips. Proteins adsorb to the hydrophobic polypropylene fibers and are eluted in a solvent suitable for MALDI‐MS analysis. Figures of merit are determined for representative compounds β2‐microglobulin, retinol binding protein, and transferrin.</p> </sec> <sec id="prca1605-sec-0030" sec-type="section"> <title>Results</title> <p>The optimum protein processing included a 100 μL aqueous rinse and an elution solvent composition was 10 μL of 55:45 ACN:water (with triflouroacetic acid). MALDI‐MS responses for the target proteins are improved from nondetectable levels to eventually yield LOD ranging from 5 to 180 nM in 1 μL aliquots.</p> </sec> <sec id="prca1605-sec-0040" sec-type="section"> <title>Conclusion and clinical relevance</title> <p>C‐CP fiber tips offer a plethora of advantages including low materials costs, high throughput, microvolume processing, and the determination of sub‐nanogram quantities of analyte; allowing determination of biomarkers that are otherwise undetectable in urine matrices.</p> </sec> </abstract>