Systematical evaluation of the effects of sample collection procedures on low-molecular-weight serum/plasma proteome profiling.
Author(s): Hsieh SY, Chen RK, Pan YH, Lee HL
Publication: Proteomics, 2006, Vol. 6, Page 3189-98
PubMed ID: 16586434 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 the effects of patient fasting, collection tube type, anticoagulant, clotting time and temperature, hemolysis, and centrifugation speed (1500 x g versus 3000 x g) and time (15 or 30 min) on serum and plasma low molecular weight proteome profiles by MALDI-TOF MS with magnetic bead fractionation. All blood was collected using a single venipuncture. Mechanical hemolysis was induced in a subset of specimens, but the authors do not describe how.
Summary of Findings:
Out of 101 defined spectral peaks, only 4 and 5 were significantly different between spectra from serum collected after an overnight fast and those from serum collected 2 and 3 h after breakfast, respectively. Out of 105 defined spectral peaks, 22 were significantly different between spectra from serum collected in plain tubes and those from serum collected in serum separator tubes (SST). With increasing clotting times at 4 degrees C or room temperature, coefficients of variation (CVs) >20% were observed in 64% and 66% of spectral peaks for serum, respectively. The authors state that increasing time before centrifugation for plasma specimens similarly increased the CV of a majority of spectral peaks. Significant differences in protein profiles were observed between plasma anticoagulated with heparin, EDTA, sodium citrate and serum. 21% of defined peaks were significantly different between serum with and without mechanically induced hemolysis, and in general, hemolysis led to additional low molecular weight peaks. Centrifugation time (15 min versus 30 min) and speed (1500 x g versus 3000 x g) had minimal effects on serum proteome profiles, with each only leading to significant differences in 4% of defined spectral peaks.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Protein MALDI-TOF MS Peptide MALDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
Biospecimen Acquisition Time of biospecimen collection Collection after overnight fast
Collection 2 h after breakfast
Collection 3 h after breakfast
Biospecimen Acquisition Type of collection container/solution Plain serum tube
SST
Biospecimen Acquisition Anticoagulant EDTA
Heparin
None
Sodium citrate
Storage Storage temperature 4 degrees C
25 degrees C
Storage Storage duration 1 h
6 h
12 h
24 h
Biospecimen Aliquots and Components Hemolysis Present
Absent
Biospecimen Aliquots and Components Centrifugation Multiple speeds compared
Multiple durations compared
Centrifugation delays investigated
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Study Purpose
The purpose of this study was to determine the effects of storage temperature and duration as well as freeze-thaw cycling of serum and plasma on low molecular weight proteome profiling by MALDI-TOF MS with magnetic bead fractionation.
Summary of Findings:
Storage of serum or plasma at 25 degrees C or 4 degrees C for up to 6 h prior to analysis led to only minimal changes in spectra, and storage for 24 h at these temperatures led to CVs of >20% for serum in 17% and 13% of spectral peaks, respectively and CVs of >20 % for plasma in 32% and 14% of spectral peaks, respectively. Storage of serum or plasma at 4 degrees C for 48 or 96 h led to a larger degree of change among spectra. Storage of serum at -80 degrees C for 3 months did not significantly affect spectral profiles nor did up to 10 freeze-thaw cycles. Freeze-thaw cycling effects were not investigated in plasma.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Protein MALDI-TOF MS Peptide MALDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
Storage Storage temperature 4 degrees C
25 degrees C
-80 degrees C
Storage Storage duration 0 h
2 h
4 h
6 h
8 h
10 h
24 h
48 h
96 h
1 month
3 months
Storage Freeze/thaw cycling 0 cycles
1 cycle
3 cycles
5 cycles
10 cycles