Franzini Raphael M, Biendl Stefan, Mikutis Gediminas, Samain Florent, Scheuermann Jörg, Neri Dario
†Institute of Pharmaceutical Sciences, ETH Zürich Vladimir-Prelog-Weg 3, 8093 Zürich, Switzerland.
‡Philochem AG, Libernstrasse 3, 8112 Otelfingen, Switzerland.
ACS Comb Sci. 2015 Jul 13;17(7):393-8. doi: 10.1021/acscombsci.5b00072. Epub 2015 Jun 17.
The potential of DNA-encoded combinatorial libraries (DECLs) as tools for hit discovery crucially relies on the availability of methods for their synthesis at acceptable purity and quality. Incomplete reactions in the presence of DNA can noticeably affect the purity of DECLs and methods to selectively remove unreacted oligonucleotide-based starting products would likely enhance the quality of DECL screening results. We describe an approach to selectively remove unreacted oligonucleotide starting products from reaction mixtures and demonstrate its applicability in the context of acylation of amino-modified DNA. Following an amide bond forming reaction, we treat unreacted amino-modified DNAs with biotinylating reagents and isolate the corresponding biotinylated oligonucleotides from the reaction mixture by affinity capture on streptavidin-coated sepharose. This approach, which yields the desired DNA-conjugate at enhanced purity, can be applied both to reactions performed in solution and to procedures in which DNA is immobilized on an anion exchange solid support.
DNA编码组合文库（DECLs）作为发现先导化合物的工具，其潜力关键取决于能否以可接受的纯度和质量合成它们的方法。在DNA存在下的不完全反应会显著影响DECLs的纯度，而选择性去除基于未反应寡核苷酸的起始产物的方法可能会提高DECL筛选结果的质量。我们描述了一种从反应混合物中选择性去除未反应寡核苷酸起始产物的方法，并证明了其在氨基修饰DNA酰化反应中的适用性。在酰胺键形成反应之后，我们用生物素化试剂处理未反应的氨基修饰DNA，并通过在链霉亲和素包被的琼脂糖上进行亲和捕获，从反应混合物中分离出相应的生物素化寡核苷酸。这种方法能以更高的纯度得到所需的DNA缀合物，可应用于溶液中进行的反应以及DNA固定在阴离子交换固体支持物上的过程。
