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Serum biomarker profiling by solid‐phase extraction with particle‐embedded micro tips and matrix‐assisted laser desorption/ionization mass spectrometry
Author(s) -
Navare Arti,
Zhou Manshui,
McDonald John,
Noriega Fernando G.,
Sullards M. Cameron,
Fernandez Facundo M.
Publication year - 2008
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3462
Subject(s) - chemistry , chromatography , mass spectrometry , matrix assisted laser desorption/ionization , surface enhanced laser desorption/ionization , sample preparation , biomarker discovery , solid phase extraction , fractionation , ion suppression in liquid chromatography–mass spectrometry , analytical chemistry (journal) , desorption , tandem mass spectrometry , proteomics , protein mass spectrometry , adsorption , biochemistry , organic chemistry , gene
One of the main challenges in high‐throughput serum profiling by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) is the development of proteome fractionation approaches that allow the acquisition of reproducible profiles with a maximum number of spectral features and minimum interferences from biological matrices. This study evaluates a new class of solid‐phase extraction (SPE) pipette tips embedded with different chromatographic media for fractionation of model protein digests and serum samples. The materials embedded include strong anion exchange (SAX), weak cation exchange (WCX), C18, C8, C4, immobilized metal affinity chromatography (IMAC) and zirconium dioxide particles. Simple and rapid serum proteome profiling protocols based on these SPE micro tips are described and tested using a variety of MALDI matrices. We show that different types of particle‐embedded SPE micro tips provide complementary information in terms of the spectral features detected for β ‐casein digests and control human serum samples. The effect of different sample pretreatments, such as serum dilution and ultrafiltration using molecular weight cut‐off membranes, and the reproducibility observed for replicate experiments, are also evaluated. The results demonstrate the usefulness of these simple SPE tips combined with offline MALDI‐TOF MS for obtaining information‐rich serum profiles, resulting in a robust, versatile and reproducible open‐source platform for serum biomarker discovery. Copyright © 2008 John Wiley & Sons, Ltd.