Low cost, high density custom peptide microarray synthesis205

描述（由申请人提供）：肽在机体的防御机制以及细胞、细菌和病毒与表面的结合中发挥重要作用。组合肽化学已经成为一种强大的工具，用于绘制药物发现应用中的受体-配体相互作用以及表位绘制。肽微阵列在药物发现、细胞通路研究和肿瘤治疗方面的应用潜力巨大，但在很大程度上尚未实现。肽微阵列尚未发挥其潜力的原因有两个：i)肽微阵列可能具有的巨大多样性以及ii)与DNA微阵列相比，肽微阵列的成本高。在本提案中，我们的目标是：1)开发一种高度灵活和快速的原位定制肽合成技术，该技术可以将肽微阵列的成本降低至少一个数量级，并将合成时间减少到少于24小时，以合成含有多达15个氨基酸的肽；2)将微阵列上多肽的密度提高一个数量级，达到100 000个/阵列；3)使用荧光探针以及高分辨率质谱来确定序列纯度和逐步添加20种天然存在的氨基酸的产量。我们开发了一种革命性的光门控寡核苷酸微阵列合成技术，该技术使用现成的试剂和改进的投影仪在开放或曲面上进行定制的微阵列合成，探针密度高达500K/玻片，成本约为大多数类似密度的商业微阵列的十分之一。在这个项目中，我们将修改用于寡核苷酸微阵列合成的化学和仪器，以开发一个在开放/关闭的载玻片表面或膜上组合肽合成的系统。

DESCRIPTION (provided by applicant): Peptides play a significant role in the defense mechanisms of the body and binding of cells, bacteria and viruses to surfaces. Combinatorial peptide chemistry has emerged as a powerful tool for mapping receptor-ligand interactions in drug discovery applications as well as epitope mapping. There is a huge, but largely unrealized, potential for peptide microarray applications in drug discovery, study of cellular pathways and treatment of tumors. There are two reasons why the peptide micro arrays have not yet reached their potential: i) the enormous diversity possible with peptide microarrays as well as ii) the high cost of peptide microarrays in comparison to DNA microarrays. In this proposal our goals are to: 1) Develop a highly flexible and fast in situ custom peptide synthesis technology which can lower the cost of peptide microarrays by at least an order of magnitude and reduce the synthesis time to less than 24 hours for peptides containing up to 15 amino acids; 2) Increase the density of peptides on a microarray by an order of magnitude to >10,000/array; 3) Use fluorescent probes as well as high resolution mass spectroscopy to determine sequence purity and stepwise yields for addition of each of the 20 naturally occurring amino acids. We have developed a revolutionary light gated oligonucleotide microarray synthesis technology which uses off the shelf reagents and a modified projector to carry out custom microarray synthesis on open or curved surfaces with probe densities of up to 500K/glass slide for about one tenth the cost of most commercial microarrays of similar density. In this project we will modify the chemistry and instrumentation used for oligonucelotide microarray synthesis to develop a system for combinatorial peptide synthesis on open/closed slide surfaces or membranes.