应用MALDI-TOF-MS技术分析HIV感染者血清多肽变化

目的用基质辅助激光解析电离飞行时间质谱（matrix-assisted laser desorption/ionization ti me offlight mass spectrometry,MALDI-TOF-MS）技术,分析艾滋病病毒（HIV）感染者和健康人血清多肽图谱的变化。方法选取20例HIV感染者、24例健康对照者的血清,经Zipplate C18层析预分离后,运用MALDI-TOF-MS分析检测,获得HIV感染者和健康人血清标本的多肽指纹图谱,并用SPSS 13.0软件对数据进行分析。随后,用质谱串联技术（MS/MS）对两个差异表达显著的多肽峰测定肽序列,在SWISS-PROT数据库进行多肽鉴定。结果在分子量为800~4 000D范围内,共发现500个多肽峰,其中11个多肽峰在HIV感染者组、正常对照组中表达均有显著差异（P〈0.05）,5个差异多肽在HIV感染者组中表达上调（m/z 1 418.67、1 465.67、1 777.82、1 864.82和2 080.23）,6个差异多肽在HIV感染者组中表达下调（m/z 904.71、920.73、1 076.73、1 521.79、1 552.75和2 080.78）;经质谱串联技术对两个表达显著差异的多肽峰测定肽序列,鉴定结果分别是纤维蛋白多肽（Fibrino-peptide）、激肽原（Kininogen processor）。结论HIV感染者和健康人血清标本的多肽指纹图谱有显著差异,通过多肽指纹图谱找到的HIV相关多肽,为今后开发HIV诊断试剂和治疗药物,提供科学依据。

Application of MALDI-TOF-MS technology for the detection of changes of serum peptides spectrum among HIV carriers

Objective To detect the spectrum of different serum peptides among HIV carriers and healthy controis by using matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI TOF MS） technology. Method MALDI TOF MS technology was used to analyze sera of 20 HIV carriers and 24 healthy controls after the sera were pre-purified by Zip Tip C18. The data were analyzed by SPSS Software 13.0. Two Peptides expressed with significant differences between HIV carriers and healthy controls were sequenced by MALDI MS/MS, and the peptides were identified via SWlSS-PROT databank. Results Five hundred peptide peaks were detected in the molecular range of 800-4000Da, among which 11 peaks were expressed with significant difference in both HIV carriers and healthy controls（P〈0.05 ）. Five of the eleven peptides（m/z 1418.67,1465.67,1777.82,1864.82 and 2080.23） were up-regulated in HIV carriers, while 6 of them （m/z 904.71,920.73,1076.73,1521.79,1552.75 and 2080. 78）were down-regulated in HIV carriers. The sequencing of 2 differently expressed peptide peaks with MS/ MS showed that they were Fibrino-peptide and Kininogen processor, respectively. Conclusion Significant differences were found in serum peptides spectrum among HIV carriers and healthy individuals, and the HIV related peptides identified via peptide spectrum may provide scientific evidence for future development of HIV diagnostic agents and therapeutic drugs.