A Novel Platform for Synthesis of Programmable Proteome-Scale Peptide Bead Arrays

用于合成可编程蛋白质组规模肽珠阵列的新型平台

基本信息

批准号：
8930164
负责人：
KURT THORN
金额：
$ 34.87万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-30 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8930164
关键词：
Allergens Allergic Allergy to peanuts Amino Acid Sequence Antibodies Antibody Repertoire Antigens Area Base Sequence Binding Biological Biological Assay Biomedical Research Code Collection DNA Microarray Chip Development Diagnostics Research Epitope Mapping Food Hypersensitivity Genome Goals Health Image Immune In Situ Length Maps Mass Spectrum Analysis Measures Medical Methods Microfluidics Nature Nucleic Acids Nucleic acid sequencing Output Patients Peptide Fragments Peptide Hydrolases Peptide Mapping Peptide Sequence Determination Peptide Synthesis Peptides Performance Phage Display Phosphoric Monoester Hydrolases Phosphotransferases Production Protein Fragment Proteins Proteome Reading Relative (related person)Ribosomes Sampling Science Serum Sorting - Cell Movement Substrate Interaction Substrate Specificity System Technology Time base biological systems cost deep sequencing enzyme substrate food allergen genome sequencing imaging system improved new technology novel novel strategies prevent protein aminoacid sequence protein protein interaction screening tool tool development

项目摘要

DESCRIPTION: Modern biomedical research has been profoundly changed by the availability of complete genome sequences and the development of tools such as deep sequencing and microarrays to leverage these sequences for both research and diagnostic applications. The availability of tools to carry out similarly large scale analyses of the whole proteome would be equally revolutionary. Such a system could be used for a broad range of applications, including mapping enzyme substrate specificity (e.g. of kinases, phosphatases, and proteases), immune-repertoire profiling (e.g. determining which specific food allergens an allergic patient has antibodies to), and protein-protein interaction screening. However, to date, the technical challenges of producing and assaying large numbers (tens of thousands) of proteins or peptides have prevented the development of such tools. Here we propose a new technology platform for the programmable synthesis and assay of proteome-scale peptide arrays. We have previously developed a microfluidic platform to produce spectrally encoded beads, which can be used as a substrate for peptide synthesis. Sorting of these beads, based on their codes, into 20 pools, will allow us to perform stepwise peptide synthesis on the beads, resulting in a collection of beads where each spectrally encoded bead is uniquely associated with a peptide sequence. This one-to-one mapping of peptides to beads allows for code-directed, step-by-step synthesis of peptides on beads and later rapid identification of bead-associated peptides by imaging alone. We expect that this technology will result in a powerful new tool for the study of antibody repertoires, enzyme-substrate interactions, and other protein-protein interactions.

描述：完整的基因组序列的可用性以及诸如深度测序和微阵列之类的工具的开发，以将这些序列用于研究和诊断应用。对整个蛋白质组进行类似大规模分析的工具的可用性将同样具有革命性。这样的系统可用于广泛的应用，包括映射酶底物特异性（例如激酶，磷酸酶和蛋白酶），免疫 - 替代性分析（例如，确定哪种特定食物过敏蛋白过敏患者具有对抗体的抗体），以及蛋白质 - 蛋白质的相互作用筛查。但是，迄今为止，生产和分析大量蛋白质或肽的技术挑战阻止了这种工具的开发。在这里，我们提出了一个新的技术平台，用于可编程合成和蛋白质尺度肽阵列的测定。我们以前已经开发了一个微流体平台来产生频谱编码的珠子，该珠子可用作肽合成的底物。这些珠子基于它们的代码将其分类为20个池，将使我们能够在珠子上执行逐步的肽合成，从而收集了一个珠子的集合，其中每个频谱编码的珠子与肽序列唯一相关。将肽对珠的一对一映射允许单独成像通过单独成像在珠子上逐步合成代码，逐步合成珠子，然后快速鉴定与珠子相关的肽。我们预计，这项技术将为研究抗体库，酶 - 基底相互作用和其他蛋白质 - 蛋白质相互作用的研究提供强大的新工具。