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Clinical Proteomics

, Volume 2, Issue 1–2, pp 67–89



Abstract

There is a great desire to relate the patterns of endogenous peptides in blood to human disease and drug response. The best practices for the preparation of blood fluids for analysis are not clear and also relatively few of the peptides in blood have been identified by tandem mass spectrometry. We compared a number of sample preparation methods to extract endogenous peptides including C18 reversed phase, precipitation, and ultrafiltration. We examined the results of these sample preparation methods by matrix-assisted laser desorption-ionization time-of-flight MALDI-TOF and liquid chromatography-tandem mass spectrometry MS-MS using MALDI-TOF-TOF and electrospray ionization-ion trap. Peptides from solid-phase extraction with C18 in the range of hundreds of femtomoles per spot were detected from the equivalent of 1 μL of serum by MALDI-TOF. We observed endogenous serum peptides from fibrinogen α- and β-chain, complement C3, α-2-HS-glycoprotein, albumin, serine or cysteine proteinase inhibitor, factor VIII, plasminogen, immunoglobulin, and other abundant blood proteins. However, we also recorded significant MS-MS spectra from tumor necrosis factor-α-, major histocompatibility complex-, and angiotensin-related peptides, as well as peptides from collagens and other low-abundance proteins. Amino acid substitutions were detected and a phosphorylated peptide was also observed. This is the first time the endogenous peptides of fetal serum have been examined by MS and where peptides from low-abundance proteins, phosphopeptides, and amino acid substitutions were detected.

Key wordsFetal calf serum liquid chromatography-electrospray ionization mass spectrometry matrix-assisted laser desorption-ionization time-of-flight MALDI ESI tandem mass spectrometry  Download to read the full article text



Autor: Declan Williams - Peihong Zhu - Peter Bowden - Catherine Stacey - Mike McDonell - Paul Kowalski - Jane Marie Kowalski - Ke

Fuente: https://link.springer.com/



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