Efficient 96-Well Plate Conjugation of Unnatural Amino Acid Building Blocks to DNA for DNA-Encoded Library Applications

Herein, we describe a robust 96-well plate workflow for high-throughput synthesis of building block-oligonucleotide conjugates for the synthesis of DNA-encoded libraries. Building blocks are Boc- and pNs-protected aminoesters, and the method allows consecutive ligation of codon oligonucleotides, ester hydrolysis, and conjugation in one pot. Results from using the two protection group strategies are discussed. Building blocks are conjugated using DMTMM to codon-specific PEG12-amino-modified oligonucleotides made from ligation. Conjugates are purified by HPLC, and all manipulations including precipitation of DNA are done in 96-well plates, reducing hands-on time.