Impact of cryopreservation on serum concentration of matrix metalloproteinases (MMP)-7, TIMP-1, vascular growth factors (VEGF) and VEGF-R2 in Biobank samples.

Author(s): Kisand K, Kerna I, Kumm J, Jonsson H, Tamm A

Publication: Clin Chem Lab Med, 2011, Vol. 49, Page 229-35

PubMed ID: 21118050 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of freeze-thaw cycling and storage temperature and duration on the stability of matrix metalloproteinases-7 (MMP-7), MMP-7 tissue inhibitors (TIMP-1), vascular endothelial growth factor (VEGF), and vascular endothelial growth factor receptor 2 (VEGF-R2) in serum.

Conclusion of Paper

TIMP-1 was relatively stable through freeze-thaw cycling and serum MMP-7 and VEGF-R2 remained stable through one freeze-thaw cycle, but three or more cycles significantly increased MMP-7 and VEGF-R2 levels. In contrast, VEGF was the least stable and decreased significantly after just one freeze-thaw cycle. MMP-7 and VEGF-R2 were relatively stable at 4 degrees C, but higher storage temperatures significantly decreased their stability. VEGF and TIMP-1 were unstable at 4 degrees C, and their instability increased with higher storage temperatures.

Studies

Study Purpose

The purpose of this study was to determine the effects of freeze-thaw cycling and storage temperature and duration on the stability of MMP-7, TIMP-1, VEGF, and VEGF-R2 in serum. Pooled serum was used for the storage temperature analysis while individual specimens were used for the freeze-thaw cycling analysis. After indicated storage treatment, all specimens were stored at -75 degrees C until analysis.

Summary of Findings:

Serum MMP-7 and VEGF-R2 remained stable through one freeze-thaw cycle, but three or more cycles increased MMP-7 levels by 24% and VEGF-R2 levels by 20%. TIMP-1 decreased by less than 13%, even after 6 freeze-thaw cycles, compared to levels in fresh serum. Serum VEGF concentrations decreased by an average of 67% after just one freeze-thaw cycle. MMP-7 was stable in serum stored for up to 35 days at 4 degrees C, but higher storage temperatures significantly decreased MMP-7 stability when compared to concentrations in serum stored at -75 degrees C. Similarly, VEGF-R2 concentrations decreased by only 12% after serum was stored for 35 days at 4 degrees C, while 21 days of storage at 40 degrees C decreased VEGF-R2 levels by 77%. VEGF and TIMP-1 were unstable at 4 degrees C, and their instability increased with higher storage temperatures. The authors extrapolate the stability of these proteins at -20 degrees C and -75 degrees C using the Arrhenius calculation.

Biospecimens

Bodily Fluid - Serum

Preservative Types

Frozen
None (Fresh)

Diagnoses:

Not specified

Platform:

Analyte	Technology Platform
Protein	ELISA

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Storage	Storage temperature	-75 degrees C 4 degrees C 20 degrees C 30 degrees C 40 degrees C
Storage	Storage duration	0 d 7 d 14 d 21 d 35 d
Storage	Freeze/thaw cycling	0 cycles 1 cycle 3 cycles 6 cycles
Biospecimen Preservation	Type of fixation/preservation	Frozen None (fresh)

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