Comparative evaluation of DNA-encoded chemical selections performed using DNA in single-stranded or double-stranded format

DNA 双绞线 计算生物学 DNA测序 生物 寡核苷酸 分子生物学 生物化学
作者
Gabriele Bassi,Nicholas Favalli,Sebastian Oehler,Adriano Martinelli,Marco Catalano,Jörg Scheuermann,Dario Neri
出处
期刊:Biochemical and Biophysical Research Communications [Elsevier]
卷期号:533 (2): 223-229 被引量:15
标识
DOI:10.1016/j.bbrc.2020.04.035
摘要

DNA-encoded chemical libraries (DEL) are increasingly being used for the discovery and optimization of small organic ligands to proteins of biological or pharmaceutical interest. The DNA fragments, that serve as amplifiable identification barcodes for individual compounds in the library, are typically used in double-stranded DNA format. To the best of our knowledge, a direct comparison of DEL selections featuring DNA in either single- or double-stranded DNA format has not yet been reported. In this article, we describe a comparative evaluation of selections with two DEL libraries (named GB-DEL and NF-DEL), based on different chemical designs and produced in both single- and double-stranded DNA format. The libraries were selected in identical conditions against multiple protein targets, revealing comparable and reproducible fingerprints for both types of DNA formats. Surprisingly, selections performed with single-stranded DNA barcodes exhibited improved enrichment factors compared to double-stranded DNA. Using high-affinity ligands to carbonic anhydrase IX as benchmarks for selection performance, we observed an improved selectivity for the NF-DEL library (on average 2-fold higher enrichment factors) in favor of single-stranded DNA. The enrichment factors were even higher for the GB-DEL selections (approximately 5-fold), compared to the same library in double-stranded DNA format. Collectively, these results indicate that DEL libraries can conveniently be synthesized and screened in both single- and double-stranded DNA format, but single-stranded DNA barcodes typically yield enhanced enrichment factors.

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