Cholesterol-Modified Oligonucleotides as Internal Reaction Controls during DNA-Encoded Chemical Library Synthesis

We report two cholesterol-modified oligonucleotides for use as internal controls for on-DNA reactions during the pooled stages of a DNA-encoded chemical library (DECL) synthesis. As these cholesterol-tagged oligonucleotides are chromatographically separable from normal DECL intermediates, they can be directly monitored by mass spectrometry to track reaction progression within a complex pool of DNA. We observed similar product conversions for reactions on substrates linked to a standard DECL DNA headpiece, to the cholesterol-modified oligonucleotides, and to the cholesterol-modified oligonucleotides while in the presence of pooled DECL synthetic intermediates.validating their use as a representative control. We also highlight an example from a DECL production in which the use of the cholesterol-modified oligonucleotides provided quality control information that guided synthetic decisions. We conclude that the use of cholesterol-modified oligonucleotides as a regular control will significantly improve the quality of DECL productions.

Automated summary

Two cholesterol-modified oligonucleotides were developed and used as internal controls in a study on DNA-encoded chemical library synthesis. These oligonucleotides, which are chromatographically separable and directly monitored by mass spectrometry, showed promising results in monitoring reaction progress and providing quality control information. The study suggests that the use of cholesterol-modified oligonucleotides as regular controls can significantly improve the quality of DNA-encoded chemical library productions.