应用表面增强激光解吸离子化飞行时间质谱筛选蕈样肉芽肿组织生物标志物

目的 采用表面增强激光解吸离子化飞行时间质谱(SELDI-TOF-MS)技术,筛选蕈样肉芽肿(MF)患者皮损组织中的生物标志物.方法 选用CMIO蛋白质芯片对12例MF患者皮损和15份正常皮肤组织提取蛋白质标本进行检测,以筛选MF患者组织中差异表达蛋白质.通过免疫沉淀方法 对部分差异表达蛋白质进行验证.结果 MF组与对照组蛋白质指纹图谱比较,发现24个差异表达蛋白质峰.将差异蛋白质峰在SWISS-PRO数据库和文献中搜索,发现相对分子质量3498的蛋白质峰与人中性粒细胞蛋白3(HNP3)相符、相对分子质量10 837的蛋白质峰与S100钙结合蛋白A8(S100A8)相符,抗S100A8特异性抗体免疫沉淀试验验证了后者.结论 SELDI-TOF-MS技术是一种快速,易行的高通量蛋白质组研究方法 ,能够鉴定出MF组织中的差异表达蛋白质,为寻找疾病相关生物学标记的提供了独特的平台.

Detection of tissue protein biomarkers by surface enhanced laser desorption/ionization-time of flightmass spectrometry in patients with mycosis fungoides

Objective To search the tissue biomarker in mycosis fungoides (MF) patients by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). Methods Skin specimens were collected from 12 MF patients and 15 normal controls to prepare solution of protein. CM10 Proteinchip was used to detect the biomarkers for MF. Differentially expressed protein was identified by immunocapturing. Results The expression intensity levels of 24 proteins were different between the MF patients and controls. By searching in SWISS-PRO database and literature, the 3498 and 10 837 peaks were found to accord with HNP3 and S100A8 proteins respectively. By immunocapturing, 10 837 could be identified as S100A8 with a special antibody against S100A8. Conclusion A quick, easy, and high throughout proteome analytic method, SELDI-TOF-MS can identify the differential expressed proteins from MF tissue, thus offering a unique platform for biomarker detection of MF.