Format

Send to

Choose Destination
Biochemistry. 2020 Jan 8. doi: 10.1021/acs.biochem.9b00919. [Epub ahead of print]

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

Author information

1
Ph.D. Programs in Chemistry and Biochemistry , CUNY Graduate Center , 365 Fifth Avenue , New York , New York 10016 , United States.
2
Firebird Biomolecular Sciences, LLC , Alachua , Florida 32615 , United States.
3
Ph.D. Program in Molecular, Cellular and Developmental Biology , CUNY Graduate Center , 365 Fifth Avenue , New York , New York 10016 , United States.
4
Department of Chemistry, Lehman College , The City University of New York , 250 Bedford Park Boulevard West , Bronx , New York 10468 , United States.
5
Foundation for Applied Molecular Evolution , 13709 Progress Boulevard , Alachua , Florida 32615 , United States.

Abstract

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.

Supplemental Content

Full text links

Icon for American Chemical Society
Loading ...
Support Center