Debaene Francois, Winssinger Nicolas
Institut de Science et Ingénierie Supramoléculaires Université Louis Pasteur-CNRS UMR 70068, Strasbourg, France.
Methods Mol Biol. 2009;570:299-307. doi: 10.1007/978-1-60327-394-7_15.
Several technologies have been described to immobilize libraries of small molecules or peptides in a microarray format. Herein, we describe protocols for an alternative strategy whereby each small molecule or peptide within a library is labeled with a peptide nucleic acid (PNA) tag such that they self-assemble in a microarray format upon hybridization with readily available DNA arrays. An important asset of the method is that it allows the library to be used in solution prior to hybridizing and as such offers the opportunity to separate the inhibitors bound to the protein from the rest of the library. Two methods based on size exclusion filtration or gel electrophoresis to separate protein-bound inhibitors from the remaining library are described.
已经有多种技术被用于以微阵列形式固定小分子或肽的文库。在此，我们描述了一种替代策略的方案，即文库中的每个小分子或肽都用肽核酸（PNA）标签进行标记，以便它们在与现成的DNA阵列杂交时以微阵列形式自组装。该方法的一个重要优点是，它允许文库在杂交之前在溶液中使用，因此提供了将与蛋白质结合的抑制剂与文库的其余部分分离的机会。本文描述了两种基于尺寸排阻过滤或凝胶电泳的方法，用于从剩余文库中分离与蛋白质结合的抑制剂。
