Solid-phase DNA-encoded library synthesis: a master builder's instructions

Area of Science:

• Medicinal Chemistry
• Chemical Biology
• Drug Discovery Technology

Background:

• DNA-encoded libraries (DELs) are a key technology in drug discovery.
• Traditional on-DNA synthesis can lead to biased binding due to DNA tag steric hindrance.
• Solid-phase DEL synthesis offers an alternative approach for lead identification.

Purpose of the Study:

• To provide a detailed protocol for solid-phase DNA-encoded library synthesis.
• To incorporate quality control innovations for robust library production.
• To enable activity-based and cellular screening of drug candidates.

Main Methods:

• On-bead synthesis of a functionalized linker using click chemistry.
• Coupling of a photocleavable linker for compound liberation.
• Combinatorial split-and-pool synthesis for library generation.
• Post-synthesis quality control via decoding, mass analysis, and deep sequencing.

Main Results:

• A robust protocol for solid-phase DEL synthesis is established.
• The method allows for physical isolation of library members.
• Quality control measures ensure library suitability for screening.
• The synthesis is accessible to molecular biology labs without extensive chemical synthesis expertise.

Conclusions:

• Solid-phase DEL synthesis is a powerful and accessible drug discovery technology.
• This method overcomes limitations of on-DNA synthesis, reducing biased binding.
• The protocol facilitates efficient lead identification through advanced screening modalities.