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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。
OBJECTIVE: To analyze the changes of serum polypeptide profiles in HIV-infected and healthy subjects by using matrix-assisted laser desorption / ionization (TI-MOMDI-TOF-MS) technique. Methods The sera of 20 HIV-infected and 24 healthy controls were selected and pre-separated by Zipplate C18. The fingerprint of the polypeptide was detected by MALDI-TOF-MS. Serum samples from HIV-infected and healthy volunteers were obtained and analyzed by SPSS 13.0 software to analyze the data. Subsequently, peptide sequences were determined by mass spectral tandem technique (MS / MS) on two polypeptide peaks differentially expressed, and peptide identification was performed on the SWISS-PROT database. Results A total of 500 peptide peaks were detected in the range of molecular weight 800-4000D. Among them, 11 polypeptide peaks were significantly different between HIV infected group and normal control group (P <0.05) The infected group was upregulated (m / z 1 418.67, 1 465.67, 1 777.82, 1 864.82 and 2 080.23), and 6 different polypeptides were downregulated in HIV infected group (m / z 904.71, 920.73, 521.79, 1 552.75 and 2 080.78). The peptide sequences were determined by tandem mass spectrometry (tandem mass spectrometry) for two polypeptide peaks showing significant differences. The identification results were Fibrino-peptide and Kininogen processor respectively. Conclusion There are significant differences in the fingerprints of human serum samples between HIV infected and healthy people. The HIV-related peptides found by peptide fingerprinting provide a scientific basis for the future development of HIV diagnostic reagents and therapeutic drugs.