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Cytokines are a group of peptides which form a sophisticated network tomodulate multiple cellular events. Within such a network, and through complexfeedback mechanisms, cytokine functions are largely interdependent and closelyassociated with a number of pathological processes. In the present study, theEVIDENCE 180 system was used to study the effects of storage temperature and repeatedfreeze/thaw cycles on the concentration of 12 cytokines in various sample types.Samples were collected from 9 healthy volunteers and stored by 3 methods: gel,glass and lithium heparin (LH) tubes. Immediately following collection, the concentrationof each cytokine in the samples was measured. Cytokine concentrations of the 3 sampletypes that did not undergo repeated freeze/thaw cycles were compared with thosesubjected to 1‑10 freeze/thaw cycles. In addition, the dynamic changes of 6 sampletypes which were stored at 4˚C for 6 h to 6 days was analyzed. In addition, thewithin‑ and between‑run precision of 12 cytokines on the biochip array was evaluated.Interleukin (IL)‑8, vascular endothelial growth factor (VEGF) and epidermal growthfactor (EGF) concentrations were lower in plasma compared with serum. Cytokinelevels in serum and plasma were affected by several freeze/thaw cycles with IL‑1β,‑4 and ‑10 increasing significantly following 1 freeze/thaw cycle and remainingat stable increased levels for the duration of the additional 9 cycles. In separatedserum samples in gel and glass tubes stored at 4˚C for 6 days, no difference inconcentration of the 12 cytokines was identified. In the other 4 sample types,IL‑8, VEGF, tumor necrosis factor α and EGF levels were altered when stored at4˚C. Results indicate that the EVIDENCE 180 system is stable and plasma was observedas the best sample type to determine concentration of the 12 cytokines using thisbiochip array. Repeated freeze/thaw cycles and storage at 4˚C was identified toaffect the concentration of the 12 cytokines. The current study demonstrates thatrepeated freeze/thaw cycles of samples must be avoid. In addition, results indicatethat plasma or serum must be separated immediately following centrifugation andsample concentration should be measured as soon as possible.

Clinical evaluation of the levels of 12 cytokines in serum/plasma under various storage conditions using evidence biochip arrays