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

描述：现代生物医学研究已经深刻地改变了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。对整个蛋白质组进行类似大规模分析的工具的可用性同样具有革命性。这样的系统可用于广泛的应用，包括绘制酶底物特异性（如激酶，磷酸酶和蛋白酶），免疫库分析（如确定过敏患者有抗体的特定食物过敏原）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（数以万计）蛋白质或肽的技术挑战阻碍了此类工具的发展。在这里，我们提出了一种新的技术平台，用于蛋白质组级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珠，它可以用作肽合成的底物。根据这些珠子的编码，将它们分成20个池，这将使我们能够对珠子进行逐步的肽合成，从而得到一组珠子，其中每个光谱编码的珠子都与肽序列唯一相关。这种一对一的多肽到微珠的映射允许在微珠上进行编码指导，一步一步合成多肽，然后通过单独成像快速识别微珠相关的多肽。我们期望这项技术将为研究抗体谱、酶-底物相互作用和其他蛋白质-蛋白质相互作用提供一个强大的新工具。

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.