Jacobs Jon M, Adkins Joshua N, Qian Wei-Jun, Liu Tao, Shen Yufeng, Camp David G, Smith Richard D
Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, PO Box 999, Richland, WA 99352, USA.
J Proteome Res. 2005 Jul-Aug;4(4):1073-85. doi: 10.1021/pr0500657.
Candidate proteomic biomarker discovery from human plasma holds both incredible clinical potential as well as significant challenges. The dynamic range of proteins within plasma is known to exceed 10(10), and many potential biomarkers are likely present at lower protein abundances. At present, proteomic based MS analyses provide a dynamic range typically not exceeding approximately 10(3) in a single spectrum, and approximately 10(4)-10(6) when combined with on-line separations (e.g., reversed-phase gradient liquid chromatography), and thus are generally insufficient for low level biomarker detection directly from human plasma. This limitation is providing an impetus for the development of experimental methodologies and strategies to increase the possible number of detections within this biofluid. Discussed is the diversity of available approaches currently used by our laboratory and others to utilize human plasma as a viable medium for biomarker discovery. Various separation, depletion, enrichment, and quantitative efforts as well as recent improvements in MS capabilities have resulted in measurable improvements in the detection and identification of lower abundance proteins (by approximately 10-10(2)). Despite these improvements, further advances are needed to provide a basis for discovery of candidate biomarkers at very low levels. Continued development of depletion and enrichment techniques, coupled with improved pre-MS separations (both at the protein and peptide level) holds promise in extending the dynamic range of proteomic analysis.
从人血浆中发现候选蛋白质组学生物标志物既具有巨大的临床潜力，也面临重大挑战。已知血浆中蛋白质的动态范围超过10的10次方，许多潜在生物标志物可能以较低的蛋白质丰度存在。目前，基于蛋白质组学的质谱分析在单光谱中提供的动态范围通常不超过约10的3次方，与在线分离（例如反相梯度液相色谱）结合时约为10的4次方至10的6次方，因此通常不足以直接从人血浆中检测低水平生物标志物。这一限制推动了实验方法和策略的发展，以增加这种生物流体中可能的检测数量。本文讨论了我们实验室和其他实验室目前使用的各种方法的多样性，这些方法将人血浆用作发现生物标志物的可行介质。各种分离、去除、富集和定量工作以及质谱能力的最新改进，已在低丰度蛋白质的检测和鉴定方面带来了可测量的改进（提高了约10至10的2次方）。尽管有这些改进，但仍需要进一步进展，为发现极低水平的候选生物标志物提供基础。去除和富集技术的持续发展，再加上质谱前分离的改进（在蛋白质和肽水平），有望扩展蛋白质组学分析的动态范围。
