Center for Preventive Doping Research/Institute of Biochemistry, German Sport University Cologne, Germany.
Proteomics. 2012 Nov;12(21):3263-72. doi: 10.1002/pmic.201200280. Epub 2012 Oct 9.
The storage of packed red blood cells (RBCs) is associated with the development of morphological and biochemical changes leading to a reduced posttransfusion functionality and viability of the cells. Within this study, 2D DIGE and high-resolution/high-accuracy Orbitrap MS were used to analyze the storage-induced changes of the cytosolic RBC proteome and identify characteristic protein patterns and potential marker proteins for the assessment of RBC storage lesions. Leukodepleted RBC concentrates were stored according to standard blood bank conditions for 0, 7, 14, 28, and 42 days and analyzed by using a characterized and validated protocol. Following statistical evaluation, a total of 14 protein spots were found to be significantly altered after 42 days of ex vivo storage. Protein identification was accomplished by tryptic digestion and LC-MS/MS and three proteins potentially useful as biomarkers for RBC aging comprising transglutaminase 2, beta actin, and copper chaperone for superoxide dismutase were selected and validated by western blot analysis. These can serve as a basis for the development of a screening assay to detect RBC storage lesions and autologous blood doping in sports.
储存浓缩红细胞（RBC）会引起形态和生化变化，从而降低细胞输注后的功能和活力。在这项研究中，使用二维 DIGE 和高分辨率/高精度 Orbitrap MS 分析了储存诱导的 RBC 胞质蛋白组变化，并鉴定了特征性蛋白质图谱和潜在的标记蛋白，用于评估 RBC 储存损伤。去白细胞的 RBC 浓缩物按照标准血库条件储存 0、7、14、28 和 42 天，并使用经过验证的方法进行分析。经过统计学评估，在体外储存 42 天后，发现共有 14 个蛋白点发生明显变化。通过胰蛋白酶消化和 LC-MS/MS 进行蛋白鉴定，选择了三个可能作为 RBC 老化生物标志物的蛋白，包括转谷氨酰胺酶 2、β肌动蛋白和超氧化物歧化酶的铜伴侣，并通过 Western blot 分析进行验证。这些可以为开发筛选检测储存损伤和运动中自体血液兴奋剂的检测提供基础。
