COMBINES-CID: An Efficient Method for De Novo Engineering of Highly Specific Chemically Induced Protein Dimerization Systems

J Am Chem Soc. 2019 Jul 17;141(28):10948-10952. doi: 10.1021/jacs.9b03522. Epub 2019 Jul 3.

Abstract

Chemically induced dimerization (CID) systems, in which two proteins dimerize only in the presence of a small molecule ligand, offer versatile tools for small molecule sensing and actuation. However, only a handful of CID systems exist and creating one with the desired sensitivity and specificity for any given ligand is an unsolved problem. Here, we developed a combinatorial binders-enabled selection of CID (COMBINES-CID) method broadly applicable to different ligands. We demonstrated a proof-of-principle by generating nanobody-based heterodimerization systems induced by cannabidiol with high ligand selectivity. We applied the CID system to a sensitive sandwich enzyme-linked immunosorbent assay-like assay of cannabidiol in body fluids with a detection limit of ∼0.25 ng/mL. COMBINES-CID provides an efficient, cost-effective solution for expanding the biosensor toolkit for small molecule detection.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques
  • Cannabidiol / analysis*
  • Dimerization
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Ligands
  • Protein Engineering*
  • Proteins / chemical synthesis*
  • Proteins / chemistry

Substances

  • Ligands
  • Proteins
  • Cannabidiol