Hartmann Daniel, Felix Klaus, Ehmann Michael, Schnölzer Martina, Fiedler Sabine, Bogumil Ralf, Büchler Markus, Friess Helmut
Research Laboratory, European Center for Pancreas Research, Section for Pancreas Surgery and Molecular Research, Department of General Surgery, University of Heidelberg, Heidelberg, Germany.
Pancreas. 2007 Nov;35(4):334-42. doi: 10.1097/mpa.0b013e3180cac723.
Testing of serum for protein patterns to monitor progression of suspected to definite chronic pancreatitis (CP).
Serum samples of CP patients and healthy volunteers were fractionated on anion exchange columns and analyzed by surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry to elucidate CP-related protein alterations and to identify biomarkers for this disease. Potential biomarkers were purified and identified by mass spectrometry.
In total, 258 protein peaks were found that discriminated between the 2 groups. Analysis revealed 28 most prominent peaks on immobilized metal affinity capture coupled with Cu and CM10 protein chips, covering the m/z range between 3.3 and 33.3 kd. Performing multivariate pattern analysis, the best pattern model was obtained using fraction 6 on immobilized metal affinity capture coupled with Cu arrays with a sensitivity of 96% and a specificity of 84%. Using a combination of matrix-assisted laser desorption-ionization-time-of-flight mass spectrometry and immunodepletion, we identified 14-m/z peaks. The proteins were found to be significantly decreased in CP serum and were identified as retinol-binding protein, serum amyloid-alpha, apolipoprotein A-II (Apo A-II), Apo C-I, Apo C-II, Apo C-III, and transthyretin and truncated forms thereof.
Distinct protein profile differences exist between normal and CP serum and reflect the metabolic and inflammatory condition in CP patients. The identified protein panel may eventually serve as a diagnostic marker set for CP.
检测血清中的蛋白质模式，以监测疑似慢性胰腺炎（CP）向确诊慢性胰腺炎的进展情况。
将CP患者和健康志愿者的血清样本在阴离子交换柱上进行分离，并用表面增强激光解吸/电离飞行时间质谱进行分析，以阐明与CP相关的蛋白质变化，并识别该疾病的生物标志物。通过质谱对潜在的生物标志物进行纯化和鉴定。
总共发现了258个可区分两组的蛋白质峰。分析显示，在固定化金属亲和捕获结合铜和CM10蛋白质芯片上有28个最突出的峰，覆盖了3.3至33.3 kd的质荷比范围。进行多变量模式分析时，使用固定化金属亲和捕获结合铜阵列上的第6部分获得了最佳模式模型，灵敏度为96%，特异性为84%。结合基质辅助激光解吸-电离飞行时间质谱和免疫耗竭技术，我们鉴定出了14个质荷比峰。这些蛋白质在CP血清中显著降低，被鉴定为视黄醇结合蛋白、血清淀粉样蛋白-α、载脂蛋白A-II（Apo A-II）、Apo C-I、Apo C-II、Apo C-III和转甲状腺素蛋白及其截短形式。
正常血清和CP血清之间存在明显的蛋白质谱差异，反映了CP患者的代谢和炎症状况。所鉴定的蛋白质组最终可能作为CP的诊断标志物集。
