质谱分析联合免疫吸附寻找系统性红斑狼疮新的血清标志物的研究

目的 探索质谱分析和免疫吸附结合来寻找系统性红斑狼疮(SLE)新的血清生物标记物的有效性.方法 采集20例活动期SLE患者(SLE组)免疫吸附(苯丙氨酸吸附柱和DNA吸附柱)前后的血清、10例非活动期SLE患者(本病对照组)、10例类风湿关节炎(RA)患者和10例干燥综合征(SS)患者(他病对照组)以及20名健康志愿者(健康对照组)的血清,与处理和活化后的壳聚糖铜衍生物纳米材料共同孵育.吸附后的纳米材料和质谱仪分析所用基质混合点板,用Axima-CFR plus基质辅助激光解析电离离子源(MALDI)飞行时间质量分析器(TOF)分析得到谱图.经软件分析找出SLE组吸附前存在而吸附后消失的峰(目标质核比峰),将该目标峰在健康对照组中进行验证,找出对照组不存在的目标质核比峰即为SLE的特异峰,再将该特异峰在其他2个对照组中进行检验.采用t检验.结果 研究共发现3个SLE特异峰蛋白,即在吸附前大量存在、在吸附后消失并且在健康对照组中不存在的质核比峰:3136、3264和3326.这些特异峰蛋白在其他2个对照组中均不存在,其相对分子质量(<10 000)均显著小于目前临床检测的SLE血清异常免疫成分如补体、抗双链DNA(dsDNA)抗体、抗Sm抗体等的相对分子质量(>50 000),因此很有可能即为本研究寻找的新的SLE血清标志物.结论 质谱分析结合免疫吸附能够有效发现新的SLE血清学标记物,有利于在健康人群中筛选鉴别出活动期SLE患者.

The detection of new serum protein biomarkers of lupus with the combination of MALDI-TOF and immunoadsorption

Objective To analyze the efficacy of the combination of MALDI-TOF and immunoadsorption to detect new biomarkers for lupus. Methods Twenty lupus patients at active stage (SLE group), 10 SLE patients in remission (SLE control group), 10 RA patients and 10 PSS patients (other rheumatic disease control group) and 20 healthy volunteers (healthy control group) were enrolled. The serum samples before and after immunoadsorption from SLE group and those from the control groups were co-incubated with activated chitosan copper derivative nano material. The adsorbed nano material was spotted onto the matrix used in MALDI-TOF for analysis by the Axima-CFR plus MALDI-TOF mass spectrometer. T-test was used for statistical analysis. Results MALDI-TOF MS screening showed that three potential protein biomarkers of mass-to-charge (m/z) ratio 3136, 3264, 3326 were found to be very specific for lupus patients: All of them were expressed before immunoadsorption in high quantity and none of them could be detected both after immunoadsorption and in all the three control groups. None of them (<10 000) were in the molecular weight range of the biomarkers used nowadays such as auto antibodies and complement (>50 000). Conclusion The combination of MALDI-TOF and immunoadsorption is effective in the detection of new serum protein biomarkers for lupus and it may be helpful in the screening of SLE patients at active stage from healthy people.