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

描述：随着全基因组序列的可获得性以及深度测序和微阵列等工具的发展，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也将是同样具有革命性的。这样的系统可用于广泛的应用，包括绘制酶底物专一性(例如，激酶、磷酸酶和蛋白水解酶)、免疫图谱分析(例如，确定过敏患者对哪些特定食物过敏原有抗体)，以及蛋白质相互作用筛选。然而，到目前为止，生产和分析大量(数万)蛋白质或多肽的技术挑战阻碍了这种工具的发展。在这里，我们提出了一个新的技术平台，用于蛋白质组规模的多肽阵列的可编程合成和分析。我们之前已经开发了一个微流控平台来生产光谱编码的珠子，它可以用作多肽合成的底物。根据编码将这些珠子分类到20个池中，将允许我们对这些珠子进行逐步的多肽合成，从而产生一个珠子集合，其中每个光谱编码的珠子都与一个肽序列唯一地关联。这种一对一的多肽到珠子的映射允许在珠子上进行代码定向、逐步合成多肽，然后仅通过成像就可以快速识别与珠子相关的多肽。我们期待这项技术将为抗体谱、酶-底物相互作用和其他蛋白质-蛋白质相互作用的研究带来一个强大的新工具。

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.