Michaud Mickael, Jourdan Eric, Villet Annick, Ravel Anne, Grosset Catherine, Peyrin Eric
Equipe de Chimie Analytique, Département de Pharmacochimie Moléculaire (UMR 5063 CNRS-UJF), ICMG FR 2607, UFR de Pharmacie de Grenoble, Université Joseph Fourier, Avenue de Verdun, 38240 Meylan, France.
J Am Chem Soc. 2003 Jul 16;125(28):8672-9. doi: 10.1021/ja034483t.
In this paper, a DNA aptamer, known to bind stereospecifically the D-enantiomer of an oligopeptide, i.e., arginine-vasopressin, was immobilized on a chromatographic support. The influence of various parameters (such as column temperature, eluent pH, and salt concentration) on the L- and D-peptide retention was investigated in order to provide information about the binding mechanism and then to define the utilization conditions of the aptamer column. The results suggest that dehydration at the binding interface, charge-charge interactions, and adaptive conformational transitions contribute to the specific D-peptide-aptamer complex formation. A very significant enantioselectivity was obtained in the optimal binding conditions, the D-peptide being strongly retained by the column while the L-peptide eluted in the void volume. A rapid baseline separation of peptide enantiomers was also achieved by modulating the elution conditions. Furthermore, it was established that the aptamer column was stable during an extended period of time. This work indicates that DNA aptamers, specifically selected against an enantiomer, could soon become very attractive as new target-specific chiral selectors for HPLC.
在本文中，一种已知能立体特异性结合寡肽（即精氨酸加压素）的D-对映体的DNA适配体被固定在色谱载体上。研究了各种参数（如柱温、洗脱液pH值和盐浓度）对L-和D-肽保留的影响，以便提供有关结合机制的信息，进而确定适配体柱的使用条件。结果表明，结合界面处的脱水、电荷-电荷相互作用以及适应性构象转变有助于形成特定的D-肽-适配体复合物。在最佳结合条件下获得了非常显著的对映体选择性，D-肽被柱强烈保留，而L-肽在空体积中洗脱。通过调节洗脱条件也实现了肽对映体的快速基线分离。此外，还确定了适配体柱在较长时间内是稳定的。这项工作表明，针对对映体特异性选择的DNA适配体可能很快成为非常有吸引力的新型高效液相色谱目标特异性手性选择剂。
