Rapid discovery of death ligands with one-bead-two-compound combinatorial library methods

Pappanaicken R. Kumaresan, Yan Wang, Mary Saunders, Yoshiko Maeda, Ruiwu Liu, Xiaobing Wang, Kit Lam

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

The one-bead-one-compound (OBOC) technology enables one to generate thousands to millions of chemical molecules on resin beads (90 μm diameter) such that each bead displays 1013 copies of the same chemical entity. Whole-cell binding assays have been developed to screen OBOC combinatorial libraries for ligands that bind to specific cell surface receptors. While very powerful, this screening method does not address the downstream cell signaling properties of the binding ligand. We have modified the OBOC technology by introducing a fixed known cell adhesion ligand to the outer layer of each bead. This one-bead-two-compound (OB2C) library configuration allows the bound cells to interact with the random immobilized chemical molecules on each bead. The bound cells can then be probed for specific cellular responses such as apoptosis and activation or inhibition of a specific cell signaling pathway. To validate this concept, an OB2C combinatorial library was created such that a random hexapeptide plus a high affinity lymphoma targeting ligand LLP2A were displayed on each bead. This LLP2A-X6 OB2C library was then screened with human T-cell leukemia cells (Molt-4) for cell death responses. After 5 days of incubation, propidium iodide was added to the bead library to stain dead cells. Beads coated by red fluorescent cells were isolated for sequence analysis. Two ligands identified by this method, when added to the lymphoid cancer cells, were able to induce cell death.

Original languageEnglish (US)
Pages (from-to)259-264
Number of pages6
JournalACS Combinatorial Science
Volume13
Issue number3
DOIs
Publication statusPublished - May 9 2011

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Keywords

  • cell signaling ligands
  • combinatorial chemistry
  • death ligands
  • high throughput screening
  • immunohistochemistry
  • one-bead-one-compound
  • one-bead-two-compound
  • peptide library

ASJC Scopus subject areas

  • Chemistry(all)

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