Processing of serum proteins underlies the mass spectral fingerprinting of myocardial infarction.
Author(s): Marshall J, Kupchak P, Zhu W, Yantha J, Vrees T, Furesz S, Jacks K, Smith C, Kireeva I, Zhang R, Takahashi M, Stanton E, Jackowski G
Publication: J Proteome Res, 2003, Vol. 2, Page 361-72
PubMed ID: 12938926 PubMed Review Paper? No
Suggested by: ISBER
Purpose of Paper
The purpose of this paper was to determine the effects of room temperature storage and addition of protease inhibitors on plasma and serum peptide profiles and to determine whether differences exist between peptide profiles of myocardial infarction patients and normal subjects.
Conclusion of Paper
Sera from myocardial infarction patients showed a distinctive peptide profile when compared to sera from normal subjects. When plasma from normal subjects was left at room temperature, a difference in peptide patterns was visible by 4 hours. Incubation of normal serum with EDTA for 4 hours had no effect on peptide patterns, but PMSF had a concentration-dependent effect. At micromolar concentrations, PMSF had little effect on the peptide pattern, but at millimolar concentrations, it reduced the signal strength of the high mass peaks and increased signal complexity in the low mass ranges. When serum from normal subjects was left at room temperature with or without PMSF, differences between the peptide patterns were noted typically after 4 to 8 hours at room temperature. With time, the addition of PMSF eliminated most of the differences in peptide profiles between sera from myocardial infarction patients and from normal subjects.
Studies
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Study Purpose
The purpose of this study was to determine whether any differences exist between serum peptide profiles of myocardial infarction and normal patients. The effects of protease inhibitors and room temperature storage of plasma on peptide profiles were also examined. All specimens, other than the fresh specimens used for room temperature storage, were stored frozen at -70 degrees C and thawed once before this study.
Summary of Findings:
Sera from myocardial infarction patients showed a distinctive peptide profile when compared to sera from normal subjects. These different profiles were seen with both MALDI-TOF and LC-ESI-ION TRAP analysis. When citrated-plasma from normal subjects was left at room temperature, a difference in peptide patterns was visible by 4 hours. Incubation of normal serum with EDTA for 4 hours had no effect on peptide patterns, but PMSF had a concentration-dependent effect. At micromolar concentrations, PMSF had little effect on the peptide pattern, but at millimolar concentrations, it reduced the signal strength of the high mass peaks and increased signal complexity in the low mass ranges. When serum from normal subjects was left at room temperature with or without PMSF, differences between the peptide patterns were noted typically after 4 to 8 hours at room temperature. With time, the addition of PMSF eliminated most of the differences in peptide profiles between sera from myocardial infarction patients and from normal subjects.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Cardiovascular Disease
- Normal
Platform:
Analyte Technology Platform Peptide MALDI-TOF MS Protein MALDI-TOF MS Peptide LC-ESI-ION TRAP Protein LC-ESI-ION TRAP Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
Analyte Extraction and Purification Protease inhibitor EDTA
PMSF
Storage Time at room temperature 0 h
4 h
8 h
24 h
Preaquisition Diagnosis/ patient condition Normal
Myocardial infarction