Hackler László, Dormán György, Kele Zoltán, Urge László, Darvas Ferenc, Puskás László G
Laboratory of Functional Genomics, Biological Research Center, Hungarian Academy of Sciences, Szeged, Temesvári krt. 62, Hungary.
Mol Divers. 2003;7(1):25-36. doi: 10.1023/b:modi.0000006534.36417.06.
Microarrays have become a widely used tool to investigate the living cell at different levels. DNA microarrays enable the expression analysis of thousand of genes simultaneously, while protein arrays investigate the properties and interactions of proteins with other proteins and with non-proteinaceous molecules. One crucial step in producing such microarrays is the permanent immobilization of samples on a solid surface. Our goal was to develop diverse linker systems capable of anchoring different biological samples, especially DNA and drug-like small molecules. We developed 6 different chemical surfaces having a 3-D-like linker system for biomolecule immobilization, and compared them to previously described immobilization strategies. The attachment chemistry utilizes the amino reactive properties of acrylic and epoxy functions. The capacity of the support was increased by creating a branching structure holding the reactive functions. The method of anchoring was investigated through a model reaction. From HPLC and mass spectrometry measurements we concluded that the covalent binding of DNA occurs through nucleobases. The tested systems offer the capability to permanently immobilize several biomolecular species in an array format.
微阵列已成为一种广泛用于在不同层面研究活细胞的工具。DNA微阵列能够同时对数千个基因进行表达分析，而蛋白质阵列则研究蛋白质与其他蛋白质以及与非蛋白质分子之间的特性和相互作用。生产此类微阵列的一个关键步骤是将样品永久固定在固体表面。我们的目标是开发出能够锚定不同生物样品，特别是DNA和类药物小分子的多种连接系统。我们开发了6种具有用于生物分子固定的三维样连接系统的不同化学表面，并将它们与先前描述的固定策略进行了比较。连接化学利用了丙烯酸和环氧官能团的氨基反应特性。通过创建一个持有反应性功能的分支结构来提高载体的容量。通过模型反应研究了固定方法。从高效液相色谱和质谱测量中我们得出结论，DNA的共价结合是通过核碱基发生的。测试的系统具备以阵列形式永久固定多种生物分子种类的能力。
