Image subtraction approach to screening one-bead-one-compound combinatorial libraries with complex protein mixtures

J Comb Chem. 2006 Jul-Aug;8(4):562-70. doi: 10.1021/cc0600268.

Abstract

To screen one-bead-one-compound (OBOC) combinatorial bead libraries,(1) one generally uses tagged purified protein as the screening probe. Compound beads that interact with the purified protein are then identified, for example, via an enzyme-linked colorimetric assay, and isolated for structure determination. In this report, we demonstrate a rapid and efficient method to screen OBOC combinatorial libraries utilizing two protein mixtures as screening probes, and by comparing optical images of the beads stained by one protein mixture but not the other, ligand beads unique to one of the two protein mixtures can be identified. The significance of this method is that it allows for rapid selection of ligands directed against proteins unique to one mixture while screening out positive beads resulting from proteins common to both mixtures as well as beads that are positive as a result of interactions with chemical and protein components found in the assay itself. The method is fast, efficient, and uses off-the-shelf equipment.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Combinatorial Chemistry Techniques*
  • Ligands
  • Microspheres*
  • Molecular Sequence Data
  • Peptide Library*
  • Protein Binding
  • Proteins / chemistry*

Substances

  • Ligands
  • Peptide Library
  • Proteins