Using hydroxymethylphenoxy derivates with the SPOT technology to generate peptides with authentic C-termini
Using hydroxymethylphenoxy derivates with the SPOT technology to generate peptides with authentic C-termini
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DOI：
10.1016/j.bmcl.2008.05.116
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发表时间：
2008-07-15
影响因子：
2.7
通讯作者：
Boisguerin, Prisca
中科院分区：
文献类型：
--
作者：
Ay, Bernhard;Landgraf, Katja;Boisguerin, Prisca
SPOT技术可以满足高平行、多肽合成可溶性多肽的要求。本文报道了一种以羟甲基苯氧乙酸（HMPA）和4-（4-羟甲基-3-甲氧基苯氧基）丁酸（HMPB）作为脯氨酸的酸性不稳定连接剂在SPOT合成中的改进方法。利用这种方法，我们可以减少通常发生在纤维素膜碱性肽裂解过程中的侧链反应。所有合成步骤都适用于全自动SPOT合成，因此代表了一个时间和成本节约的程序。此外，改进的切割和洗涤步骤产生了具有真实c -末端的肽，纯度在60-95%之间。我们改进的方法非常适合于合成数千种不同的肽，然后直接用于需要真实c -末端的不同生物测定，如CD8 t细胞表位筛选，疫苗免疫或肿瘤成像。(c) 2008 Elsevier Ltd.版权所有。
The SPOT technology can fulfill most requirements for highly parallel, multiple peptide synthesis of soluble peptides within the upper microgram range. Here, we report on an improved method using hydroxymethylphenoxyacetic acid (HMPA) for 19 amino acids and 4-(4-hydroxymethyl-3-methoxyphenoxy)-butyric acid (HMPB) for proline as acidic labile linkers in SPOT synthesis. Using this approach we could reduce side-chain reactions normally occurring during conventional alkaline peptide cleavage from cellulose membranes. All synthesis steps were adapted to fully-automated SPOT synthesis and therefore represent a time- and cost-saving procedure. Furthermore, the improved cleavage and washing steps resulted in peptides with authentic C-termini in a purity range of 60-95%. Our improved method is ideal for synthesizing many thousand different peptides subsequently used directly for different biological assays requiring authentic C-termini, such as CD8 T-cell epitope screening, vaccine immunization, or tumor imaging. (c) 2008 Elsevier Ltd. All rights reserved.