Lee Dong Hyeon, Kim Jun Woo, Jeon Soon Young, Park Bo Kyung, Han Bok Ghee
Department of Physiology, School of Medicine, CHA University, Gyeonggi-do, Korea.
Ann Clin Lab Sci. 2010 Winter;40(1):61-70.
This study examined the effect of storage temperature on the protein profile of human serum. Serum samples were stored for > or =7 days at -80 degrees C, -20 degrees C, 4 degrees C, and room temperature prior to proteomic analysis. Serum protein fractionations by SDS-PAGE, including high and low molecular weight (MW) proteins, showed that several bands had different intensities after storage at higher temperatures. Fractionations by 2-dimensional gel electrophoresis (2-DE) indicated that approximately 60 protein spots had changed significantly after storage at higher temperatures. These proteins included C3/C4, alpha2-macroglobulin, and alpha1B glycoprotein, based on identification by MS and MS/MS. The profile of the low MW serum proteins, analyzed using SELDI ProteinChip Arrays (IMAC3 and Q10), was also significantly changed after storage at higher temperatures. These results indicate that the higher storage temperatures have a significant influence on serum protein profiles regardless of MW. Based on these findings, serum specimens should be preferably be stored at -80 degrees C before proteomic analysis.
本研究检测了储存温度对人血清蛋白质谱的影响。在进行蛋白质组分析之前，血清样本分别在-80℃、-20℃、4℃和室温下储存≥7天。通过SDS-PAGE对血清蛋白进行分级分离，包括高分子量和低分子量蛋白，结果显示在较高温度下储存后，几条条带的强度有所不同。二维凝胶电泳（2-DE）分级分离表明，在较高温度下储存后，约60个蛋白点发生了显著变化。基于质谱（MS）和串联质谱（MS/MS）鉴定，这些蛋白包括C3/C4、α2-巨球蛋白和α1B糖蛋白。使用表面增强激光解吸电离飞行时间质谱（SELDI）蛋白质芯片阵列（IMAC3和Q10）分析的低分子量血清蛋白谱，在较高温度下储存后也发生了显著变化。这些结果表明，无论分子量大小，较高的储存温度对血清蛋白质谱都有显著影响。基于这些发现，在进行蛋白质组分析之前，血清标本最好储存在-80℃。
