Photochemical patterning of biological molecules inside a glass capillary
Photochemical patterning of biological molecules inside a glass capillary
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DOI：
10.1021/ac0504619
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发表时间：
2005-09-01
影响因子：
7.4
通讯作者：
Chatelain, F
中科院分区：
文献类型：
--
作者：
Balakirev, MY;Porte, S;Chatelain, F
本文介绍了一种将生物分子光化学图案化到熔融石英毛细管内壁上的简单方法。该方法是基于与光活性二苯甲酮（BP）衍生物的内毛细管表面的改性。在365 nm处对填充有样品溶液的毛细管进行UV照射，导致溶质通过稳定的共价键与BP部分交联。作为概念的证明，使用倒置显微镜作为照射装置将寡核苷酸和蛋白质排列在毛细管内。我们证明了以这种方式产生的毛细管阵列是功能性的，并且可以用于不同的生物测定，包括DNA杂交、蛋白质相互作用研究和免疫测定。毛细管阵列具有与平面形式的基于荧光团的测定相当的灵敏度，具有几个优点，包括亚微升样品体积和短的测定时间。因此，在许多低密度阵列应用中，如突变检测和诊断免疫测定中，毛细管格式应被视为平面格式的可能替代品。
A simple way for photochemical patterning of biological molecules onto the inner wall of fused-silica capillary is described. The method is based on a modification of the inner capillary surface with photoactive benzophenone (BP) derivative. The UV irradiation at 365 nm of the capillary filled with a sample solution results in cross-linking of the solutes to the BP moiety via a stable covalent bond. As a proof of concept, oligonucleotides and proteins were arrayed inside the capillary using an inverted microscope as an irradiation device. We demonstrated that the capillary arrays produced in this way are functional and could be used in different bioassays including DNA hybridization, protein interaction studies, and immunoassays. Having a sensitivity comparable to the fluorophore-based assays in a planar format, the capillary array possesses several advantages including submicroliter sample volume and a short assay time. The capillary format should therefore be considered as a possible alternative to a planar format in a number of low-density array applications such as mutation detection and diagnostic immunoassays.