DETECTION OF INTACT PEPTIDES BY MATRIX ASSISTED TOF SSIMS: MASS SPECT

通过基质辅助 TOF SSIMS 检测完整肽：质谱

• 批准号: 6345030
• 负责人: PETER KINGSHOTT
• 金额: $ 1.84万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6345030
• 关键词: animal tissue; bioengineering /biomedical engineering; biological products; biomaterials; biomedical equipment development; biomedical resource; biotechnology; proteins; technology /technique

In this project the Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI) concept is being extended for use in Time-of-Flight Static Secondary Ion Mass Spectrometry (ToF-SSIMS). The overall aim of the project is to use MALDI matrix molecules to increase both the ion yields of intact proteins and peptides and to reduce fragmentation caused by Cs+ ion beam impact that occurs in standard SIMS analysis. Initial work is directed towards the detection of intact model peptide molecules. Model peptides with ?-sheet (YGLKLKLKL-NH2 Mw=1075.42 amu) and ?-helix (YG(LKKLLKL)2-NH2 Mw=1911.34 amu)) tertiary structures (synthesized using standard FMOC solid phase synthesis) are being used in conjunction with standard MALDI matrices [2,5-dihydroxybenzoic acid (DHB) and 3,5-dimethoxy-4-hydroxycinnamic (sinapinic acid (SA))] to facilitate generation of molecular ions upon irradiation with the SIMS Cs+ ion >source. SIMS imaging with micrometer spatial resolution of mat rix/peptid es crystals on Si wafers has to date suggested that this mode of analysis can be applied to specifically map the distribution of peptide molecules within MALDI crystals.

在该项目中，基质辅助激光解吸电离 质谱 (MALDI) 概念正在扩展用于 飞行时间静态二次离子质谱 (ToF-SSIMS)。 该项目的总体目标是利用 MALDI 基质分子 增加完整蛋白质和肽的离子产率，并 减少因 Cs+ 离子束撞击而产生的碎片 标准 SIMS 分析。 初步工作的目标是 完整模型肽分子的检测。 模型肽 β-折叠 (YGLKLKLKL-NH2 Mw=1075.42 amu) 和 β-螺旋 (YG(LKKLLKL)2-NH2 Mw=1911.34 amu)) 三级结构（使用标准 FMOC 合成 固相合成）与标准结合使用 MALDI 基质 [2,5-二羟基苯甲酸 (DHB) 和 3,5-二甲氧基-4-羟基肉桂酸（芥子酸（SA））]促进 SIMS Cs+ 离子照射后产生分子离子 >来源。 具有微米空间分辨率的垫子 SIMS 成像 迄今为止，Si 晶片上的 rix/peptid es 晶体表明，这 分析模式可用于具体绘制分布图 MALDI 晶体内的肽分子。