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一种用于蛋白质微阵列的新型聚合物涂层。

A new polymeric coating for protein microarrays.

作者信息

Cretich Marina, Pirri Giovanna, Damin Francesco, Solinas Isabella, Chiari Marcella

机构信息

Istituto di Chimica del Riconoscimento Molecolare (ICRM), C.N.R., Via Mario Bianco 9, 20131 Milano, Italy.

出版信息

Anal Biochem. 2004 Sep 1;332(1):67-74. doi: 10.1016/j.ab.2004.05.041.

Abstract

Despite the increasing interest in arraying proteins in a high-density format, several technical issues still impede the development of protein microarray technology. One of the major problems is the availability of substrates that are able to bind native proteins with high density. In this study, we investigated the suitability of a novel surface as a support for protein microarrays. A polymeric glass coating is obtained by physical adsorption of a N,N-dimethylacrylamide (DMA), N,N-acryloyloxysuccinimide (NAS), and [3-(methacryloyl-oxy)propyl]trimethoxysilyl (MAPS) copolymer. The coating procedure provides a fast and inexpensive method of producing hydrophilic functional surfaces. The slide performance was investigated in a protein-protein interaction experiment and in the assessment of rheumatoid factor (RF) in human serum samples. The results demonstrate that the ligands immobilized on the polymeric surface maintain an active conformation and are easily accessible, providing a detection limit of 54amol/spot. Moreover, in the RF assay, after hybridization with the sera, the slides have a low background, leading to a detection limit of 900amol/spot.

摘要

尽管人们对以高密度形式排列蛋白质的兴趣日益浓厚,但一些技术问题仍然阻碍着蛋白质微阵列技术的发展。主要问题之一是缺乏能够高密度结合天然蛋白质的底物。在本研究中,我们研究了一种新型表面作为蛋白质微阵列支撑物的适用性。通过物理吸附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)共聚物获得聚合物玻璃涂层。该涂层工艺提供了一种快速且廉价的制备亲水性功能表面的方法。在蛋白质 - 蛋白质相互作用实验以及人血清样本中类风湿因子(RF)的评估中对载玻片性能进行了研究。结果表明,固定在聚合物表面的配体保持活性构象且易于接近,检测限为54 amol/斑点。此外,在RF检测中,与血清杂交后,载玻片背景较低,检测限为900 amol/斑点。

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