Coelho Graça Didia, Hartmer Ralf, Jabs Wolfgang, Beris Photis, Clerici Lorella, Stoermer Carsten, Samii Kaveh, Hochstrasser Denis, Tsybin Yury O, Scherl Alexander, Lescuyer Pierre
Department of Human Protein Sciences, Faculty of Medicine, Geneva University, Rue Michel Servet 1, 1211, Geneva, Switzerland.
Anal Bioanal Chem. 2015 Apr;407(10):2837-45. doi: 10.1007/s00216-015-8525-5. Epub 2015 Mar 10.
Hemoglobin disorder diagnosis is a complex procedure combining several analytical steps. Due to the lack of specificity of the currently used protein analysis methods, the identification of uncommon hemoglobin variants (proteoforms) can become a hard task to accomplish. The aim of this work was to develop a mass spectrometry-based approach to quickly identify mutated protein sequences within globin chain variants. To reach this goal, a top-down electron transfer dissociation mass spectrometry method was developed for hemoglobin β chain analysis. A diagnostic product ion list was established with a color code strategy allowing to quickly and specifically localize a mutation in the hemoglobin β chain sequence. The method was applied to the analysis of rare hemoglobin β chain variants and an (A)γ-β fusion protein. The results showed that the developed data analysis process allows fast and reliable interpretation of top-down electron transfer dissociation mass spectra by nonexpert users in the clinical area.
血红蛋白疾病诊断是一个结合多个分析步骤的复杂过程。由于目前使用的蛋白质分析方法缺乏特异性，鉴定罕见的血红蛋白变体（蛋白质异构体）可能成为一项难以完成的任务。这项工作的目的是开发一种基于质谱的方法，以快速鉴定珠蛋白链变体中的突变蛋白序列。为实现这一目标，开发了一种自上而下的电子转移解离质谱方法用于血红蛋白β链分析。通过颜色编码策略建立了一个诊断产物离子列表，可快速、特异性地定位血红蛋白β链序列中的突变。该方法应用于分析罕见的血红蛋白β链变体和一种（A）γ-β融合蛋白。结果表明，所开发的数据分析过程使临床领域的非专业用户能够快速、可靠地解释自上而下的电子转移解离质谱图。
