HUPO Plasma Proteome Project specimen collection and handling: Towards the standardization of parameters for plasma proteome samples
Author(s): Rai Alex J, Gelfand Craig A, Haywood Bruce C, Warunek David J, Yi Jizu, Schuchard Mark D, Mehigh Richard J, Cockrill Steven L, Scott Graham BI, Tammen Harald, Schulz-Knappe Peter, Speicher David W, Vitzthum Frank, Haab Brian B, Siest Gerard, Chan Daniel W
Publication: Proteomics, 2005, Vol. 5, Page 3262
PubMed ID: 16052621 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine if peptide content differs in paired plasma and serum samples via RP-HPLC MALDI-TOF mass spectrometry.
Summary of Findings:
Of the peptide signals present in serum, only 40% were also identified in plasma. Peptides differentially expressed in serum ranged from 1-5 kD and included intracellular, coagulation dependent, and enzymatically derived peptides.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Peptide MALDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
-
Study Purpose
To determine if peptide content differs as a function of storage duration and temperature.
Summary of Findings:
While the peptide profile of serum samples stored at -20, -80 degrees C, or in liquid nitrogen for 2 months did not differ significantly, multiple differences were observed in peptide content and expression in samples stored at 4 or 23 degrees C after 30 days. Enzymatic activity and protein stability were also affected in several examined proteins: lactate dehydrogenase, aspartate-amino transferase, alkaline phosphatase, LDL, HDL.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Protein 1D/2D gels Peptide SELDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage temperature Liquid nitrogen
-80 degrees C
-20 degrees C
4 degrees C
23 degrees C
Storage Storage duration 0 hr
2 hr
4 hr
8 hr
1 d
2 d
7 d
30 d
60 d
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Study Purpose
To determine if peptide content and expression differs as a result of incubation with a protease inhibitor cocktail and its individual components.
Summary of Findings:
Plasma samples incubated with a protease inhibitor cocktail differed significantly from controls and displayed higher signals. Protease inhibitor treated plasma that was albumin and IgG depleted exhibited an altered native pI profile for several proteins, and general smearing. Examination of protease inhibitor cocktail components indicated that 4-(2-aminoethyl) benzene sulfonyl fluoride (AEBSF) was responsible for adverse effects.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Peptide SELDI-TOF MS Protein 1D/2D gels Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Protease inhibitor E-64
4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF)
aprotinin
leupeptin
bestatin
pepstatin A
Cocktail
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Study Purpose
To determine whether the background signal of antibody based microarrays are influenced by protease inhibitor treatment at various times during sample preparation.
Summary of Findings:
Addition of a protease inhibitor cocktail both before and after labeling for antibody microarray analysis decreased background and improved signal to background ratio.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Protein Antibody microarray Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Analyte Extraction and Purification Protease inhibitor Added during analysis
Added prior to analysis
Added prior to and during analysis