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

点击翻译按钮获取中文摘要

*** 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. ***