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

• 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
• 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

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.

产品说明：现代生物医学研究已经被完全基因组序列的可用性和工具的发展，如深度测序和微阵列，以利用这些序列的研究和诊断应用的深刻变化。对整个蛋白质组进行类似的大规模分析的工具的可用性同样具有革命性。这样的系统可以用于广泛的应用，包括映射酶底物特异性（例如激酶、磷酸酶和蛋白酶的）、免疫谱分析（例如确定过敏患者对哪些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，生产和测定大量（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，我们提出了一个新的技术平台，可编程的蛋白质组规模的肽阵列的合成和测定。我们之前已经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。基于它们的代码，将这些珠子分选到20个池中，将允许我们在珠子上进行逐步肽合成，从而产生珠子的集合，其中每个光谱编码的珠子与肽序列独特地相关联。这种肽与珠子的一对一映射允许在珠子上编码指导的逐步合成肽，并且随后仅通过成像快速鉴定珠子相关的肽。我们预计，这项技术将导致一个强大的新工具，用于抗体库，酶-底物相互作用，和其他蛋白质-蛋白质相互作用的研究。