Time-Dependent and Sample-to-Sample Variations in Human Plasma Peptidome are Both Minimized Through Use of Protease Inhibitors
Author(s): Craft D, Yi Jizu, Gelfand CA
Publication: Analytical Letters, 2009, Vol. 42, Page 1398-1406
Review Paper? No
Purpose of Paper
The purpose of this paper was to compare the plasma peptidome between EDTA blood collected into tubes with and without protease inhibitors both when blood was processed immediately and after a processing delay.
Conclusion of Paper
While plasma peptide content was greater when blood was collected into EDTA tubes that lacked protease inhibitors, EDTA plasma from collection tubes that contained a protease inhibitor cocktail displayed more uniform peptide spectra between individuals and more stable peptide content after a 4 hour processing delay.
Studies
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Study Purpose
The purpose of this study was to determine whether blood collected into tubes containing protease inhibitors display an altered EDTA plasma peptidome and to determine the effect of protease inhibitors on between-patient variability. Blood was collected from three healthy individuals into two tubes: EDTA tubes, P100 tubes (contains EDTA and a protease inhibitor cocktail). Blood was left in the original collection tube at room temperature for 0, 60, 120, 180, or 240 min prior to plasma processing by centrifugation and filtration. Peptide spectra were visualized after fractionation on a nano-HPLC system.
Summary of Findings:
Blood collected in tubes containing a protease inhibitor cocktail (P100 tubes), displayed more uniform peptide spectra following room temperature storage compared to those collected into EDTA tubes. Variability among peptide spectra between patients was also reduced among specimens collected in P100 tubes in comparison to EDTA tubes.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Peptide HPLC Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 0 min
60 min
120 min
180 min
240 min
Analyte Extraction and Purification Protease inhibitor No protease inhibitor added
Cocktail
Biospecimen Acquisition Type of collection container/solution BD Vacutainer PPT (EDTA)
BD P100 (EDTA and Protease inhibitor cocktail)
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Study Purpose
The purpose of this study was to determine whether blood collected into tubes containing protease inhibitors display an altered plasma peptidome, compared to EDTA plasma specimens, immediately or after a 240 min processing delay. Blood was collected from a single healthy individual into two tubes: EDTA, P100 tubes (contains EDTA and a protease inhibitor cocktail). Blood was left in the original collection tube at room temperature for 0 or 240 min prior to plasma processing by centrifugation and filtration. Peptides extracted from plasma were analyzed by LC-MALDI-MS.
Summary of Findings:
Peptide content was greater in EDTA tubes compared to P100 tubes when plasma was processed immediately. However, when tubes were subjected to a 4 hour processing delay peptide content decreased considerable in EDTA tubes but remained stable in P100 tubes. Differences between tube types that were processed immediately suggest that proteolysis occurs rapidly in the absence of protease inhibitors.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Peptide MALDI-TOF MS Peptide HPLC Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 0 min
240 min
Analyte Extraction and Purification Protease inhibitor No protease inhibitor added
Cocktail
Biospecimen Acquisition Type of collection container/solution BD Vacutainer PPT (EDTA)
BD P100 (EDTA and Protease inhibitor cocktail)