Proteomic analysis of the effect of storage temperature on human serum.

Author(s): Lee DH, Kim JW, Jeon SY, Park BK, Han BG

Publication: Ann Clin Lab Sci, 2010, Vol. 40, Page 61-70

PubMed ID: 20124332 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of storage temperature on serum protein profiles of specimens stored for more than 7 d. Individual specimens were analyzed for all experiments except SELDI-TOF mass spectrometry (MS) where pooled serum from 8 donors was analyzed.

   Summary of Findings:

   Several changes were observed in SDS-PAGE gels after storage of serum at temperatures above -80 degrees C consistent with protein degradation or modification. The changes observed were more evident in major protein depleted serum compared to whole serum. A significant increase was seen in the number of spots on 2-DE gels for serum stored at room temperature (923) compared to numbers observed after serum was stored at -80, -20, or 4 degrees C (646, 625, and 626, respectively). Spots appeared, disappeared, increased in intensity or decreased when results for different storage temperatures were compared. Specifically, changes were seen in 29 spots with increasing storage temperatures and 33 spots only showed changes when serum was stored at room temperature compared to other storage temperatures. The authors identified these protein spots through MALDI-TOF MS or MS/MS and included proteins such as C3/C4, alpha2-macroglobulin, and alpha1B glycoprotein. SELDI ProteinChip analysis detected much higher peak signals from serum stored at 4 degrees C as opposed to -80 degrees C. The IMAC3-Zinc (Zn) array detected 80 peaks, and 12.5% significantly changed between the two storage temperatures. The IMAC3-Copper (Cu) array detected 23 peaks, and 30.4% of these significantly changed between storage temperatures. The Q10 array detected 47 peaks, and 6.4% of these significantly changed between storage temperatures.

   Biospecimens

   • Bodily Fluid - Serum

   Preservative Types

   • None (Fresh)
   • Other Preservative
   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	1D/2D gels
   Protein	MS/MS
   Protein	MALDI-TOF MS
   Protein	SELDI-TOF MS
   Peptide	1D/2D gels
   Peptide	MALDI-TOF MS
   Peptide	SELDI-TOF MS
   Peptide	MS/MS

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage temperature	Room temperature 4 degrees C -20 degrees C -80 degrees C
   SELDI-TOF MS Specific	Type of array	IMAC3-Zn IMAC3-Cu Q10
   Biospecimen Aliquots and Components	Biospecimen components	Whole serum Major protein depleted serum
   Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh) Refrigeration

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