Normalization of DNA encoded library affinity selection results driven by high throughput sequencing and HPLC...

Area of Science:

• Drug Discovery
• Medicinal Chemistry
• Biotechnology

Background:

• Affinity selection screening generates vast data for identifying target-enriched compounds.
• The analytical approach is critical for successful DNA Encoded Library (DEL) platforms.
• Current methods can be biased by chemical reaction yields, affecting compound selection.

Purpose of the Study:

• To present a combined purification and sequencing method for DEL library production.
• To improve the quality of DEL libraries before affinity selection.
• To enable normalization of affinity selection data, reducing bias from chemical yields.

Main Methods:

• Integrating High Performance Liquid Chromatography (HPLC) purification into DEL library production.
• Utilizing Next Generation Sequencing (NGS) analysis to assess pre-selection reaction yields.
• Applying normalization analysis to affinity selection output data.

Main Results:

• Achieved higher quality DEL libraries through integrated HPLC purification.
• Enabled accurate assessment of chemical reaction yields using NGS prior to selection.
• Minimized bias in affinity selection analysis by normalizing for reaction yield.

Conclusions:

• The combined HPLC and NGS approach significantly enhances DEL library quality.
• This method provides a robust framework for reliable affinity selection data analysis.
• It facilitates more accurate short-listing of compounds for synthesis and drug development.