Chersi A, Sezzi M L, Romano T F, Evangelista M, Nista A
Regina Elena Institute for Cancer Research, Rome, Italy.
Biochim Biophys Acta. 1990 Jun 20;1034(3):333-6. doi: 10.1016/0304-4165(90)90060-a.
In this study, several methods for controlled labelling of synthetic peptides by the use of fluorescent compounds (fluorescein isothiocyanate and dimethylaminonaphthalene sulfonyl chloride) were investigated. The first reagent yielded monofluoresceinated, active compounds only when the peptides lacked lysine residues. Monolabelling of peptides in solution with dimethylaminonaphthalenesulphonyl chloride was hindered by the broad reactivity of the reagent, but was achieved by reacting the fluorochrome on protected resin-bound peptides in solid-phase synthesis. The remarkable stability of the linkage allowed the cleavage of the peptide from the resin and deprotection of side-chain functions without hydrolysis of the labelled group. The binding of antipeptide antibodies to the labelled fragments was then estimated using different techniques.
在本研究中，对几种使用荧光化合物（异硫氰酸荧光素和二甲基氨基萘磺酰氯）对合成肽进行可控标记的方法进行了研究。第一种试剂仅在肽缺乏赖氨酸残基时产生单荧光素化的活性化合物。用二甲基氨基萘磺酰氯对溶液中的肽进行单标记受到该试剂广泛反应性的阻碍，但通过在固相合成中使荧光染料与受保护的树脂结合肽反应得以实现。连接的显著稳定性使得肽能够从树脂上裂解下来并对侧链功能进行脱保护，而不会使标记基团水解。然后使用不同技术评估抗肽抗体与标记片段的结合情况。
