Screening of a one bead-one compound combinatorial library for β-actin identifies molecules active toward Ramos B-lymphoma cells

Suzanne Miyamoto, Ruiwu Liu, Susan Hung, Xiaobing Wang, Kit Lam

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The search for small molecules that specifically recognize protein targets is a laborious process if conducted in a one protein-one compound manner. A high-throughput antibody-based screening of one bead-one compound (OBOC) combinatorial small molecule libraries is described here, whereby libraries containing thousands of different small molecule ligands are synthesized on individual TentaGel beads and simultaneously screened for protein binding to individual beads, each with a different compound. The use of OBOC libraries greatly facilitates this simultaneous screening of thousands of compounds. Now, through the use of monoclonal or affinity-purified antibodies, small molecules that bind a particular protein contained in a complex mixture of biological molecules have been identified. This method identified small molecule ligands that bound β-actin present in cytoplasmic cell extracts of Ramos B-lymphoma cells. These small molecule ligands were resynthesized in immobilized and soluble forms and tested for binding of β-actin present in Ramos B-cell extracts and for activity against Ramos lymphoma cells. This high-throughput screening immunoassay method has great promise for improving our ability to find relevant, bioactive small molecules that target a specific native protein in a complex protein mixture without purification of the protein.

Original languageEnglish (US)
Pages (from-to)112-120
Number of pages9
JournalAnalytical Biochemistry
Volume374
Issue number1
DOIs
StatePublished - Mar 1 2008

Fingerprint

B-Cell Lymphoma
Actins
Screening
Molecules
Proteins
Ligands
Cell Extracts
Complex Mixtures
Libraries
High-Throughput Screening Assays
Small Molecule Libraries
Antibody Affinity
Throughput
Immunoassay
Protein Binding
Antibodies
Lymphoma
B-Lymphocytes
Purification
Cells

Keywords

  • Antibody
  • Combinatorial library
  • High throughput screening
  • Immunoassay
  • Proliferation
  • Ramos B-lymphoma
  • Small molecule

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Screening of a one bead-one compound combinatorial library for β-actin identifies molecules active toward Ramos B-lymphoma cells. / Miyamoto, Suzanne; Liu, Ruiwu; Hung, Susan; Wang, Xiaobing; Lam, Kit.

In: Analytical Biochemistry, Vol. 374, No. 1, 01.03.2008, p. 112-120.

Research output: Contribution to journalArticle

Miyamoto, Suzanne ; Liu, Ruiwu ; Hung, Susan ; Wang, Xiaobing ; Lam, Kit. / Screening of a one bead-one compound combinatorial library for β-actin identifies molecules active toward Ramos B-lymphoma cells. In: Analytical Biochemistry. 2008 ; Vol. 374, No. 1. pp. 112-120.
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