Instituto de Biomedicina de Málaga (IBIMA)
A sustainable alternative to deep-freezing for plasma and serum preservation.
Plasma is a complex biological fluid with a high protein content and significant clinical relevance. It is particularly sensitive to freeze-thaw cycles and storage conditions, which can affect the stability of its components and compromise downstream analyses.
Objectives
Comparison of values obtained by quantitative determination of the biomarkers leptin, adiponectin and PCR in serum samples collected and frozen at -80ºC and serum samples subjected to a drying procedure and stored at room temperature. Samples will be collected from obese donors and healthy controls.
Materials and Methods
Samples used for the study
Plsma and serum samples
60 samples
Fresh
Procedure
Serum: Centrifuge at 1600g for 10 minutes at room temperature. Collect and aliquot into 500 µL volumes for freezing at -80°C and lyophilization.
Plasma and buffy coat: Centrifuge at 1,500g for 10 minutes. Separate plasma and buffy coat into 500 µL aliquots and store half at -80°C and half for lyophilization.
Sampling times will be fresh, 1 and 6 months.
Quality Assurance
ELISA
ELISAs will be performed to detect leptin, adiponectin and CRP, proteins commonly altered in patients with obesity.
Both CRP and adiponectin are biomarkers of cardiovascular risk, while leptin is considered important in the development of obesity because it affects both energy intake and energy expenditure by participating in the satiety signaling pathway in the brain. In addition, the adiponectin/leptin ratio is used to study this pathology, so they are related biomarkers. These three markers are altered in patients with obesity with respect to the control population, making their determination optimal for validating this technique in a real research environment.
Results
Coming soon…
Conclusion
Coming soon…
