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 TAR这个小型企业创新研究第一阶段项目将探索多肽和蛋白质测序与高通量质谱学相结合的可行性，使用一种类似于埃德曼化学的新方法。初步实验表明，其他人试图使传统的埃德曼降解法适应于产生用于质谱分析的截断集(“梯子”)而产生的几个问题，将被这种基于二硫化碳的替代化学所绕过。将讨论关于氨基酸残基和不同多肽的物理/化学特性的一般性问题。将建立和优化反应条件、控制、一致性(重复性、可预测性)和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和序列信息的研究人员是可取的，以识别新的蛋白质，确认从DNA预测的序列，定位翻译后或实验修饰的位置，分析生物活性多肽，将2D凝胶上观察到的斑点连接到DNA数据库中的信息(蛋白质组研究)，并提供重组蛋白质的质量控制。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. ***