Ligand-Guided Selection with Artificially Expanded Genetic Information Systems against TCR-CD3ε

Biochemistry. 2020 Feb 4;59(4):552-562. doi: 10.1021/acs.biochem.9b00919. Epub 2020 Jan 8.


Here we are reporting, for the first time, a ligand-guided selection (LIGS) experiment using an artificially expanded genetic information system (AEGIS) to successfully identify an AEGIS-DNA aptamer against T cell receptor-CD3ε expressed on Jurkat.E6 cells. Thus, we have effectively combined the enhanced diversity of an AEGIS DNA library with LIGS to develop a superior screening platform to discover superior aptamers. Libraries of DNA molecules from highly diversified building blocks will provide better ligands due to more functional diversity and better-controlled folding. Thus, a DNA library with AEGIS components (dZ and dP) was used in LIGS experiments against TCR-CD3ε in its native state using two clinically relevant monoclonal antibodies to identify an aptamer termed JZPO-10, with nanomolar affinity. Multiple specificity assays using knockout cells, and competition experiments using monoclonal antibodies utilized in LIGS, show unprecedented specificity of JZPO-10, suggesting that the combination of LIGS with AEGIS-DNA libraries will provide a superior screening platform to discover artificial ligands against critical cellular targets.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aptamers, Nucleotide / genetics
  • Aptamers, Nucleotide / immunology
  • CD3 Complex / genetics*
  • CD3 Complex / immunology*
  • CD3 Complex / metabolism
  • Gene Library
  • Humans
  • Jurkat Cells
  • Ligands
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / immunology
  • SELEX Aptamer Technique / methods


  • Aptamers, Nucleotide
  • CD3 Complex
  • CD3E protein, human
  • Ligands
  • Receptors, Antigen, T-Cell