scholarly journals Evaluation of Sample Preparation Methods for Fast Proteotyping of Microorganisms by Tandem Mass Spectrometry

2019 ◽  
Vol 10 ◽  
Author(s):  
Karim Hayoun ◽  
Duarte Gouveia ◽  
Lucia Grenga ◽  
Olivier Pible ◽  
Jean Armengaud ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Sample Preparation ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Preparation Methods ◽  
Sample Preparation Methods

scholarly journals Comparison of methods to examine the endogenous peptides of fetal calf serum

2021 ◽  
Author(s):  
Declan Williams ◽  
Peihong Zhu ◽  
Peter Bowden ◽  
Catherine Stacey ◽  
Mike McDonell ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Amino Acid ◽  
Sample Preparation ◽  
Tandem Mass Spectrometry ◽  
Amino Acid Substitutions ◽  
Tandem Mass ◽  
Preparation Methods ◽  
Endogenous Peptides ◽  
Maldi Tof ◽  
Sample Preparation Methods

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.

scholarly journals Comparison of methods to examine the endogenous peptides of fetal calf serum

2021 ◽  
Author(s):  
Declan Williams ◽  
Peihong Zhu ◽  
Peter Bowden ◽  
Catherine Stacey ◽  
Mike McDonell ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Amino Acid ◽  
Sample Preparation ◽  
Tandem Mass Spectrometry ◽  
Amino Acid Substitutions ◽  
Tandem Mass ◽  
Preparation Methods ◽  
Endogenous Peptides ◽  
Maldi Tof ◽  
Sample Preparation Methods

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.

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