Immobilized OBOC combinatorial bead array to facilitate multiplicative screening

Wenwu Xiao, Fernanda C. Bononi, Jared Townsend, Yuanpei Li, Ruiwu Liu, Kit Lam

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

One-bead-one-compound (OBOC) combinatorial library screening has been broadly utilized for the last two decades to identify small molecules, peptides or peptidomimetics targeting variable screening probes such as cell surface receptors, bacteria, protein kinases, phosphatases, proteases etc. In previous screening methods, library beads were suspended in solution and screened against one single probe. Only the positive beads were tracked and isolated for additional screens and finally selected for chemical decoding. During this process, the remaining negative beads were not tracked and discarded. Here we report a novel bead immobilization method such that a bead library array can be conveniently prepared and screened in its entirety, sequentially many times with a series of distinct probes. This method not only allows us to increase the screening efficiency but also permits us to determine the binding profile of each and every library bead against a large number of target receptors. As proof of concept, we serially screened a random OBOC disulfide containing cyclic heptapeptide library with three water soluble dyes as model probes: malachite green, bromocresol purple and indigo carmine. This multiplicative screening approach resulted in a rapid determination of the binding profile of each and every bead respective to each of the three dyes. Beads that interacted with malachite green only, bromocresol purple only, or both indigo carmine and bromocresol purple were isolated, and their peptide sequences were determined with microsequencer. Ultimately, the novel OBOC multiplicative screening approach could play a key role in the enhancement of existing on-bead assays such as whole cell binding, bacteria binding, protein binding, posttranslational modifications etc. with increased efficiency, capacity, and specificity.

Original languageEnglish (US)
Pages (from-to)441-448
Number of pages8
JournalCombinatorial Chemistry and High Throughput Screening
Volume16
Issue number6
DOIs
StatePublished - Jul 2013

Keywords

  • High throughput screening
  • Multiplicative screening
  • One-bead-one-compound combinatorial chemistry
  • PDMS affixed bead array
  • Water soluble organic dye

ASJC Scopus subject areas

  • Organic Chemistry
  • Drug Discovery
  • Computer Science Applications

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