Dental Research Institute, University of California, Los Angeles, CA 90095, USA.
Anal Chim Acta. 2012 Apr 13;722:63-9. doi: 10.1016/j.aca.2012.02.017. Epub 2012 Feb 19.
Human saliva is a biological fluid with emerging early detection and diagnostic potentials. However, the salivary proteome suffers from rapid degradation and thus compromises its translational and clinical utilities. Therefore, easy, reliable and practical methods are urgently required for the storage of human saliva samples. In this study, saliva samples from healthy subjects were collected and stored at room temperature (RT) and 4 °C for different lengths of time with and without specific protein stabilization treatments. SDS-PAGE was run to compare the protein profiling between samples. Reference proteins, β-actin and interleukin-1 β (IL1β), were chosen to evaluate salivary protein stability. Immunoassay was used for the detection of these target proteins. All data was compared with the positive control that had been kept at -80 °C. The results show that the salivary proteome that has been stored at 4 °C with added protease inhibitors was stable for approximately two weeks without significant degradation. By adding ethanol to the samples, the salivary proteome was stabilized at RT. After optimization, a simple, robust and convenient method is developed for the stabilization of proteins in human saliva that does not affect the downstream translational and clinical applications. The salivary proteome could be stabilized without significant degradation by adding ethanol at RT for about two weeks. This optimized method could greatly accelerate the clinical usage of saliva for future diagnosis.
唾液是一种具有早期检测和诊断潜力的生物液体。然而，唾液蛋白质组易迅速降解，从而影响其转化和临床应用。因此，急需简单、可靠且实用的方法来储存人唾液样本。在这项研究中，收集了健康受试者的唾液样本，并在室温（RT）和 4°C 下，分别在有和没有特定蛋白质稳定处理的情况下，储存不同的时间。运行 SDS-PAGE 以比较样本之间的蛋白质图谱。选择β-肌动蛋白和白细胞介素-1β（IL1β）作为参考蛋白来评估唾液蛋白质的稳定性。免疫测定法用于检测这些靶蛋白。所有数据均与保存在-80°C 的阳性对照进行比较。结果表明，在添加蛋白酶抑制剂的情况下，4°C 储存的唾液蛋白质组在大约两周内没有明显降解，是稳定的。通过向样本中添加乙醇，可在 RT 下稳定唾液蛋白质组。经过优化，开发了一种简单、稳健且方便的方法，可稳定人唾液中的蛋白质，而不会影响下游的转化和临床应用。通过在 RT 下添加乙醇，唾液蛋白质组可以在大约两周内保持稳定，而不会出现明显降解。这种优化方法可以极大地加速唾液在未来诊断中的临床应用。
