Two-dimensional gel electrophoresis of serum specimens from a normal population.

Author(s): Tracy RP, Currie RM, Young DS

Publication: Clin Chem, 1982, Vol. 28, Page 890-9

PubMed ID: 6176364 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 blood collection method, anticoagulant, addition of protease inhibitor, delayed analysis, and frozen storage on the electrophoretic patterns of plasma and serum from healthy individuals. Albumin and immunoglobulin G (IgG) were removed from specimens prior to assay, and gels were silver stained.

   Summary of Findings:

   Heparinized plasma contained alpha, beta and gamma bands of fibrinogen and prothrombin bands. In serum, fibrinogen bands were absent, and prothrombin bands were mostly absent, but two unidentified bands were observed that were not observed in plasma. When plasma specimens were stored for 4-6 months, at -20 degrees C, prior to analysis, two reproducible spots appeared that were not present in fresh plasma. The larger of these spots was not observed in frozen serum, but the smaller was observed after frozen storage of plasma or serum. The authors report that using capillary rather than venous plasma, a 2 h room temperature delay in processing and using EDTA instead of heparin as an anticoagulant had no effects on electrophoretic patterns. The addition of PMSF in ethanol to whole blood prior to plasma preparation resulted in the presence of erythrocyte lysate proteins that were not generally observed in plasma prepared without PMSF, and the authors attributed these proteins to the addition of ethanol. The standard electrophoretic patterns created using 2-dimensional electrophoresis very strongly correlated with those previously determined using radial immunodiffusion.

   Biospecimens

   • Bodily Fluid - Serum
   • Bodily Fluid - Plasma
   • Bodily Fluid - Blood

   Preservative Types

   • None (Fresh)
   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Glycoprotein	Radial immunodiffusion assay
   Protein	1D/2D gels
   Protein	Radial immunodiffusion assay
   Glycoprotein	1D/2D gels

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh)
   Storage	Time at room temperature	0 h 2 h
   Storage	Storage duration	0 h 4-6 months
   Biospecimen Aliquots and Components	Blood and blood products	Plasma Serum
   Biospecimen Acquisition	Method of fluid acquisition	Finger/heel prick sampling Venipuncture
   Biospecimen Acquisition	Anticoagulant	EDTA Heparin None
   Analyte Extraction and Purification	Protease inhibitor	No protease inhibitor added PMSF
   1D/2D gels Specific	Technology platform	Radial immunodiffusion
   Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated
   Biospecimen Acquisition	Anatomical location of blood draw	Capillary Vein

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