Serum peptide profiling by magnetic particle-assisted, automated sample processing and MALDI-TOF mass spectrometry Journal Article
| Authors: | Villanueva, J.; Philip, J.; Entenberg, D.; Chaparro, C. A.; Tanwar, M. K.; Holland, E. C.; Tempst, P. |
| Article Title: | Serum peptide profiling by magnetic particle-assisted, automated sample processing and MALDI-TOF mass spectrometry |
| Abstract: | Human serum contains a complex array of proteolytically derived peptides (serum peptidome) that may provide a correlate of biological events occurring in the entire organism; for instance, as a diagnostic for solid tumors (Petricoin, E. F.; Ardekani, A. M.; Hitt, B. A.; Levine, P. J.; Fusaro, V. A.; Steinberg, S. M.; Mills, G. B.; Simone, C.; Fishman, D. A.; Kohn, E. C.; Liotta, L. Lancet 2002, 359, 572-577). Here, we describe a novel, automated technology platform for the simultaneous measurement of serum peptides that is simple, scalable, and generates highly reproducible patterns. Peptides are captured and concentrated using reversed-phase (RP) batch processing in a magnetic particle-based format, automated on a liquid handling robot, and followed by a MALDI TOF mass spectrometric readout. The protocol is based on a detailed investigation of serum handling, RP ligand and eluant selection, small-volume robotics design, an optimized spectral acquisition program, and consistent peak extraction plus binning across a study set. The improved sensitivity and resolution allowed detection of 400 polypeptides (0.8-15-kDa range) in a single droplet (∼50 μL) of serum, and almost 2000 unique peptides in larger sample sets, which can then be analyzed using common microarray data analysis software. A pilot study indicated that sera from brain tumor patients can be distinguished from controls based on a pattern of 274 peptide masses. This, in turn, served to create a learning algorithm that correctly predicted 96.4% of the samples as either normal or diseased. |
| Keywords: | brain neoplasms; sensitivity analysis; mass spectrometry; protein blood level; blood proteins; tumor markers, biological; cancer screening; algorithms; automation; pilot projects; glioblastoma; peptides; robotics; image processing, computer-assisted; matrix assisted laser desorption ionization time of flight mass spectrometry; plasma protein; complexation; magnetics; spectrometry, mass, matrix-assisted laser desorption-ionization; polypeptides; peptide analysis; autoanalysis; data acquisition; coated materials, biocompatible; humans; article; learning algorithms; eluants |
| Journal Title: | Analytical Chemistry |
| Volume: | 76 |
| Issue: | 6 |
| ISSN: | 0003-2700 |
| Publisher: | American Chemical Society |
| Date Published: | 2004-03-15 |
| Start Page: | 1560 |
| End Page: | 1570 |
| Language: | English |
| DOI: | 10.1021/ac0352171 |
| PROVIDER: | scopus |
| PUBMED: | 15018552 |
| DOI/URL: | |
| Notes: | Anal. Chem. -- Cited By (since 1996):356 -- Export Date: 16 June 2014 -- CODEN: ANCHA -- Source: Scopus |
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