Cretich Marina, Longhi Renato, Corti Angelo, Damin Francesco, Di Carlo Gabriele, Sedini Valentina, Chiari Marcella
Istituto di Chimica del Riconoscimento Molecolare (ICRM)-C.N.R. Milano, Milano, Italy.
Methods Mol Biol. 2009;570:221-32. doi: 10.1007/978-1-60327-394-7_10.
In this chapter we report on the characterization of linear antigenic sites of human chromogranin A (CgA), a useful tissue and serum marker for neuroendocrine tumours and a precursor of many biologically active peptides. The epitope mapping of CgA has been carried out by peptide microarrays on glass slides coated by a copolymer of N,N-dimethylacrylamide (DMA), N,N-acryloyloxysuccinimide (NAS) and [3-(methacryloyl-oxy) propyl] trimethoxysilyl (MAPS). The microarray support provided sufficient accessibility of the ligand, with no need for a spacer, as the polymer chains prevent interaction of immobilized peptides with substrate. In addition, the polymeric surface constitutes an aqueous micro-environment in which, despite peptide random orientation, linear epitopes are freely exposed. The results reported are in accordance with those obtained in conventional ELISA assays using biotinylated and non-biotinylated peptides.
在本章中，我们报告了人嗜铬粒蛋白A（CgA）线性抗原表位的特征，CgA是神经内分泌肿瘤的一种有用的组织和血清标志物，也是许多生物活性肽的前体。CgA的表位图谱是通过肽微阵列在由N，N - 二甲基丙烯酰胺（DMA）、N，N - 丙烯酰氧基琥珀酰亚胺（NAS）和[3 - （甲基丙烯酰氧基）丙基]三甲氧基硅烷（MAPS）的共聚物包被的载玻片上进行的。该微阵列载体为配体提供了足够的可及性，无需间隔物，因为聚合物链可防止固定化肽与底物相互作用。此外，聚合物表面构成了一个水性微环境，在其中，尽管肽呈随机取向，但线性表位仍可自由暴露。报告的结果与使用生物素化和非生物素化肽的传统ELISA分析中获得的结果一致。
