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Design, Synthesis, and Application of OB2C Combinatorial Peptide and Peptidomimetic Libraries

  • Ruiwu Liu
  • Tsung-Chieh Shih
  • Xiaojun Deng
  • Lara Anwar
  • Sara Ahadi
  • Pappanaicken Kumaresan
  • Kit S. Lam
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1248)

Abstract

The “one-bead two-compound” (OB2C) combinatorial library is constructed on topologically segregated trifunctional bilayer beads such that each bead has a fixed cell-capturing ligand and a random library compound co-displayed on its surface and a chemical coding tag (bar code) inside the bead. An OB2C library containing thousands to millions of compounds can be synthesized and screened concurrently within a short period of time. When live cells are incubated with such OB2C libraries, every bead will be coated with a monolayer of cells. The cell membranes of the captured cells facing the bead surface are exposed to the library compounds tethered to each bead. A specific biochemical or cellular response can be detected with an appropriate reporter system. The OB2C method enables investigators to rapidly discover synthetic molecules that not only interact with cell-surface receptors but can also stimulate or inhibit downstream cell signaling. To demonstrate this powerful method, one OB2C peptide library and two OB2C peptidomimetic libraries were synthesized and screened against Molt-4 lymphoma cells to discover “death ligands.” Apoptosis of the bead-bound cells was detected with immunocytochemistry using horseradish peroxidase (HRP)-conjugated anti-cleaved caspase-3 antibody and 3,3′-diaminobenzidine as a substrate. Two novel synthetic “death ligands” against Molt-4 cells were discovered using this OB2C library approach.

Key words

OB2C combinatorial library Apoptosis Immunocytochemistry Cell-capturing ligand Synthetic “death ligand” Bilayer bead Proapoptotic agent Lymphoma 

Notes

Acknowledgments

We would like to thank Michael Schlein for editorial assistance. This work was supported by NIH R33CA160132 and institutional fund from UC Davis.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Ruiwu Liu
    • 1
  • Tsung-Chieh Shih
    • 1
  • Xiaojun Deng
    • 1
  • Lara Anwar
    • 1
  • Sara Ahadi
    • 1
  • Pappanaicken Kumaresan
    • 2
  • Kit S. Lam
    • 1
  1. 1.Department of Biochemistry & Molecular MedicineUniversity of California DavisSacramentoUSA
  2. 2.Department of PediatricsUniversity of Texas MD Anderson Cancer CenterSacramentoUSA

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