Profiling post-centrifugation delay of serum and plasma with antibody bead arrays.
Author(s): Qundos U, Hong MG, Tybring G, Divers M, Odeberg J, Uhlen M, Nilsson P, Schwenk JM
Publication: J Proteomics, 2013, Vol. 95, Page 46-54
PubMed ID: 23631827 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of plasma collection tube type and post-centrifugation storage of plasma and serum in the collection tube on the protein profile using custom antibody bead arrays.
Conclusion of Paper
For all specimen types, the inter-individual variability was greater than the intra-individual variability across storage durations. Plasma obtained using either plasma preparation tubes (PPT) or standard EDTA tubes had comparable intensity distributions which were wider and lower than those found for serum. Although 86% (322 of 373) of antibodies gave higher signals for serum than plasma collected in either type of tube, the biggest differences (3-4 fold) were in proteins that had higher expression in plasma such as fibrinogen, Proto-oncogene tyrosine-protein kinase receptor Ret (Ret), and Carbohydrate sulfotransferase 8 (CHST8). Caldesmon 1 binding was higher when plasma was collected in standard EDTA tubes than PPT tubes and declined significantly with storage of plasma in standard EDTA tubes, but not PPT tubes. While binding of 20 of the 373 antibodies tested showed inter-individual variation during storage, other than caldesmon 1 binding levels, none of the variation was found to be significant.
Studies
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Study Purpose
The purpose of this study was to determine the effects of plasma collection tube type and post-centrifugation storage of plasma and serum in the collection tube on the protein profile using custom antibody bead arrays. Blood from 16 healthy patients was drawn into 2 EDTA-plasma preparation tubes (PPT), 2 EDTA tubes without gel separator, and 2 serum separator tubes. Plasma was centrifuged immediately and serum was centrifuged within 40 min. Plasma and serum tubes were stored at both 4°C and 23°C in the original collection containers. At each time-point, an aliquot was removed from the tube and frozen at -80°C until analysis using 373 antibodies.
Summary of Findings:
For all specimen types, the inter-individual variability was greater than the intra-individual variability across storage durations. Plasma obtained using PPT and standard EDTA tubes had comparable intensity distributions which were wider and lower than those found for serum. 86% (322 of 373) of antibodies gave higher signals for serum than plasma collected in either type of tube, but the biggest differences (3-4 fold) were in proteins that had higher expression in plasma and included fibrinogen (3.5-3.6-fold, q<2E-27 for PPT, q<5.0E-28 for standard EDTA), Ret (3.3-fold, q=4.0E-29 for PPT and q=5.3E-30 for standard EDTA), and CHST8 (3.2-fold, q=5.1E-29 for PPT and q=3.7E-30 for standard EDTA). Importantly, comparison of the protein profile in PPT plasma and standard EDTA plasma revealed that caldesmon 1 was higher when plasma was collected in standard EDTA tubes than PPT tubes. Caldesmon 1 significantly decreased (slope=-0.14, false discovery rate, FDR=3E-03) with storage of plasma in standard EDTA tubes, regardless of storage temperature. However in PPT tubes, this decrease was not significant. While binding of 20 of the 373 antibodies tested showed inter-individual variation with storage, none was found to be significant.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform Protein Antibody microarray Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution PPT
Standard EDTA tube
SST
Biospecimen Aliquots and Components Blood and blood products Plasma
Serum
Storage Storage duration 1 h
3 h
8 h
36 h
Storage Storage temperature 4°C
23°C