NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis
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A novel multiplex-protein array for serum diagnostics of colorectal cancer: impact of pre-analytical storage conditions.

Author(s): Bünger S, Klempt-Giessing K, Toner V, Kelly M, FitzGerald SP, Brenner H, von Eggeling F, Habermann JK

Publication: Biopreserv Biobank, 2013, Vol. 11, Page 379-86

PubMed ID: 24835368 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of frozen storage temperature and up to 3 freeze-thaw cycles on the levels of 8 proteins in serum from patients with colorectal cancer.

Conclusion of Paper

Storing serum at -80 rather than -170 degrees C had no significant effects on levels of the 8 tested proteins, but non-significant decreases in carcinoembryonic antigen (CEA) with storage at -80 degrees C were large enough to lead to a false negative malignancy result for 1 of 3 patients. In contrast, while freeze-thaw cycling had no diagnostic impact, it led to significantly increased levels of CEA, Interleukin-8 (IL-8), complement component 3 a (C3adesArg) and C-reactive protein (CRP) and decreased levels of vascular endothelial growth factor (VEGF). Macrophage colony-stimulating factor (M-CSF) levels were lower after 2 freeze-thaw cycles than 1, but after 3 freeze-thaw cycles, M-CSF levels were comparable to those measured after 1 freeze-thaw cycle. Calgizzarin (S100A1) and CD26 were not affected by freeze-thaw cycling of serum.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of frozen storage temperature and up to 3 freeze-thaw cycles on the levels of 8 proteins in serum from patients with colorectal cancer. Serum specimens were collected from 20 patients during post-operative follow-ups. Serum was stored, within 30 min of collection, at both -80 and -170 degrees C. Specimens were stored for 22.2-35.7 months before thawing on ice and analysis. Specimens that had been stored at -170 degrees C were returned to liquid nitrogen, thawed and assayed again 24 and 48 h later. Protein levels were determined using a multiplex chemiluminescent sandwich immunoassay biochip.

    Summary of Findings:

    Nonsignificant decreases in the levels of CD26 and CRP, and nonsignificant increases in IL-8, VEGF, S100A11, and C3adesArg were observed when serum was stored at -80 degrees C rather than at -170 degrees C. The decrease in CEA levels with storage at -80 degrees C was large enough to lead to a false negative malignancy result for 1 of 3 patients. In contrast, while freeze-thaw cycling had no diagnostic impact, it led to significantly increased levels of CEA (p<0.001), IL-8 (p<0.001), C3adesArg (p<0.001) and CRP (p=0.043) and significantly decreased levels of VEGF (p<0.0001). M-CSF levels were lower after 2 freeze-thaw cycles than 1 (p=0.002), but after 3 freeze-thaw cycles, M-CSF levels were comparable to those measured after 1 freeze-thaw cycle. S100A1 and CD26 levels were not affected by freeze-thaw cycling of serum.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Neoplastic - Carcinoma
    Platform:
    AnalyteTechnology Platform
    Protein Immunoassay
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Freeze/thaw cycling 1 cycle
    2 cycles
    3 cycles
    Storage Storage temperature -80 degrees C
    -170 degrees C
    View more

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