SBIR Phase I: Use of Carbon Disulfide to Generate Peptide Sequencing Ladders for Analysis by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

* 9660710塔尔 这个小企业创新研究第一阶段项目将探索肽和蛋白质测序与高通量质谱联用的可行性，使用类似于埃德曼化学的新方法。 初步实验表明，由其他人尝试调整常规Edman降解以产生用于质谱分析的截短集（“梯”）而产生的几个问题将通过基于二硫化碳的这种替代化学来规避。 关于氨基酸残基和不同肽的物理/化学特性的一般性问题将得到解决。 将建立和优化反应条件、控制、一致性（可重复性、可预测性）以及后续自动化的充分基础。 能够实现这种化学的试剂盒对于使用肽图谱和序列信息来鉴定新蛋白质、确认从DNA预测的序列、定位翻译后或实验修饰的位点、分析生物活性肽、将在2D凝胶上观察到的点与DNA数据库中的信息联系起来（“蛋白质组”研究）以及提供重组蛋白质的质量控制的研究人员来说是期望的。 第二阶段的自动化仪器将与传统的埃德曼测序仪竞争。 ***

*** 9660710 Tarr This Small Business Innovation Research Phase I project will explore the feasibility of peptide and protein sequencing coupled to high throughput mass spectrometry using a novel methodology resembling the Edman chemistry. Preliminary experiments suggest that the several problems arising from the attempts by others to adapt the conventional Edman degradation to produce truncation sets ("ladders") for mass spectral analysis will be circumvented by this alternative chemistry based on carbon disulfide. Issues of generality with respect to amino acid residues and to the physical/chemical characteristics of different peptides will be addressed. Reaction conditions, control, consistency (repeatability, predictability), and adequate basis for later automation will be established and optimized. Kits enabling this chemistry would be desirable to researchers using peptide maps and sequence information to identify novel proteins, confirm sequences predicted from the DNA, locate sites of post-translational or experimental modifications, analyze bioactive peptides, tie spots observed on 2D gels to information in DNA databases ("proteome" research), and provide quality control of recombinant proteins. An automated instrument from Phase II would compete well with conventional Edman sequencers. ***