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：
8762296
负责人：
KURT THORN
金额：
$ 48.88万
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/8762296
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 Image Imaging Device Immune In Situ Length Maps Mass Spectrum Analysis Measures Medical Methods Microfluidics Nature Nucleic Acids Nucleic acid sequencing Output Patients Peanuts - dietary 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 improved new technology novel novel strategies prevent protein aminoacid sequence protein protein interaction public health relevance 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：由于完整基因组序列的可用性以及深度测序和微阵列等工具的开发，现代生物医学研究已经发生了深刻的变化，以利用这些序列进行研究和诊断应用。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，包括绘制酶底物特异性（例如激酶、磷酸酶和蛋白酶的）、免疫库分析（例如确定过敏患者对哪种特定食物过敏原具有抗体）和蛋白质-蛋白质相互作用筛选。然而，迄今为止，生产和分析大量（数万）蛋白质或肽的技术挑战阻碍了此类工具的开发。在这里，我们提出了一种用于蛋白质组规模肽阵列的可编程合成和测定的新技术平台。我们之前开发了一个微流体平台来生产光谱编码的珠子，它可以用作肽合成的底物。根据这些珠子的代码将它们分类为 20 个池，这将使我们能够在珠子上进行逐步肽合成，从而形成一个珠子集合，其中每个光谱编码的珠子都与肽序列唯一相关。这种肽与珠子的一对一映射允许在珠子上进行肽的代码引导、逐步合成，并随后仅通过成像快速识别珠子相关的肽。我们预计这项技术将成为研究抗体库、酶-底物相互作用和其他蛋白质-蛋白质相互作用的强大新工具。