Serum Peptide Profiling by Magnetic Particle-Assisted, Automated Sample Processing and MALDI-TOF Mass Spectrometry Export

@article{citeulike:4159291, abstract = {PMID: 15018552 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, 572577). 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 (0.815-kDa range) in a single droplet (50 muL) 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.}, author = {Villanueva, Josep and Philip, John and Entenberg, David and Chaparro, Carlos A. and Tanwar, Meena K. and Holland, Eric C. and Tempst, Paul}, citeulike-article-id = {4159291}, citeulike-linkout-0 = {http://dx.doi.org/10.1021/ac0352171}, citeulike-linkout-1 = {http://pubs.acs.org/doi/abs/10.1021/ac0352171}, day = {1}, doi = {10.1021/ac0352171}, journal = {Analytical Chemistry}, keywords = {maldi, serum}, month = {March}, number = {6}, pages = {1560--1570}, posted-at = {2009-11-05 14:22:18}, priority = {2}, title = {Serum Peptide Profiling by Magnetic Particle-Assisted, Automated Sample Processing and MALDI-TOF Mass Spectrometry}, url = {http://dx.doi.org/10.1021/ac0352171}, volume = {76}, year = {2004} }

TY - JOUR ID - citeulike:4159291 L3 - citeulike-article-id:4159291 N2 - PMID: 15018552 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, 572577). 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 (0.815-kDa range) in a single droplet (50 muL) 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. IS - 6 JF - Analytical Chemistry EP - 1570 TI - Serum Peptide Profiling by Magnetic Particle-Assisted, Automated Sample Processing and MALDI-TOF Mass Spectrometry VL - 76 SP - 1560 KW - maldi KW - serum AU - Villanueva, Josep AU - Philip, John AU - Entenberg, David AU - Chaparro, Carlos AU - Tanwar, Meena AU - Holland, Eric AU - Tempst, Paul PY - 2004/03/01/ UR - http://dx.doi.org/10.1021/ac0352171 ER -