High-throughput screening of one-bead-one-compound peptide libraries using intact cells

Choi Fong Cho, Babak Behnam Azad, Leonard G. Luyt, John D. Lewis

Research output: Contribution to journalArticle

Abstract

Screening approaches based on one-bead-one-compound (OBOC) combinatorial libraries have facilitated the discovery of novel peptide ligands for cellular targeting in cancer and other diseases. Recognition of cell surface proteins is optimally achieved using live cells, yet screening intact cell populations is time-consuming and inefficient. Here, we evaluate the Complex Object Parametric Analyzer and Sorter (COPAS) large particle biosorter for high-throughput sorting of bead-bound human cell populations. When a library of RGD-containing peptides was screened against human cancer cells that express αvβ 3 integrin, it was found that bead-associated cells are rapidly dissociated when sorted through the COPAS instrument. When the bound cells were reversibly cross-linked onto the beads, however, we demonstrated that cell/bead mixtures can be sorted quickly and accurately. This reversible cross-linking approach is compatible with matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based peptide sequence deconvolution. This approach should allow one to rapidly screen an OBOC library and identify novel peptide ligands against cell surface targets in their native conformation.

Original languageEnglish (US)
Pages (from-to)393-400
Number of pages8
JournalACS Combinatorial Science
Volume15
Issue number8
DOIs
Publication statusPublished - Aug 12 2013
Externally publishedYes

    Fingerprint

Keywords

  • cell-based screen
  • COPAS biosorter
  • cross-linking
  • integrin
  • MALDI-TOF mass spectrometry
  • OBOC library
  • RGD peptide

ASJC Scopus subject areas

  • Chemistry(all)

Cite this