Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion
Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion
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DOI：
10.1038/nprot.2007.160
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Hancock, Robert E. W.
中科院分区：
文献类型：
--
作者：
Hilpert, Kai;Winkler, Dirk F. H.;Hancock, Robert E. W.
利用SPOT技术在纤维素上合成多肽，可以以少量平行合成大量可寻址的多肽。此外，每个肽的成本不到传统树脂合成肽的1%。SPOT方法遵循传统纤维素片上的标准氟酰-甲氧基-羰基化学反应，可以使用600多种不同的构建模块。该过程包括三个阶段：纤维素膜的制备，氨基酸的逐步偶联和侧链保护基团的裂解。如有必要，肽可以从膜上切割出来，用可溶性肽进行测定。这些特点使该方法成为筛选大量多肽用于许多不同目的的极好工具。潜在的应用范围从简单的结合分析到更复杂的酶分析和活微生物或细胞的研究。完成程序所需的时间取决于肽的数量和长度。例如，可以在6天内合成400个9-mer肽。
Peptide synthesis on cellulose using SPOT technology allows the parallel synthesis of large numbers of addressable peptides in small amounts. In addition, the cost per peptide is less than 1% of peptides synthesized conventionally on resin. The SPOT method follows standard fluorenyl-methoxy-carbonyl chemistry on conventional cellulose sheets, and can utilize more than 600 different building blocks. The procedure involves three phases: preparation of the cellulose membrane, stepwise coupling of the amino acids and cleavage of the side-chain protection groups. If necessary, peptides can be cleaved from the membrane for assays performed using soluble peptides. These features make this method an excellent tool for screening large numbers of peptides for many different purposes. Potential applications range from simple binding assays, to more sophisticated enzyme assays and studies with living microbes or cells. The time required to complete the protocol depends on the number and length of the peptides. For example, 400 9-mer peptides can be synthesized within 6 days.