鈥淐ap-and-Catch鈥?Purification for Enhancing the Quality of Libraries of DNA Conjugates
详细信息 查看全文
- 作者：Raphael M. Franzini ; Stefan Biendl ; Gediminas Mikutis ; Florent Samain ; J枚rg Scheuermann ; Dario Neri
- 刊名：ACS Combinatorial Science
- 出版年：2015
- 出版时间：July 13, 2015
- 年：2015
- 卷：17
- 期：7
- 页码：393-398
- 全文大小：310K
- ISSN：2156-8944
文摘
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.