title: Optimization of the Magnetic Recovery of Hits from One-Bead-One-Compound Library Screens.
abstract: On-bead screening of one-bead-one-compound (OBOC) libraries is a useful procedure for the identification of protein ligands. An important aspect of this experiment is the method by which beads that bind the target protein are separated from those that do not. Ideally, such a method would be rapid and convenient and result in the isolation of 100% of the "hits" with no false positives (beads that display compounds that are not good ligands for the target). We introduced a technique in which beads that have bound a labeled target protein can be magnetized, thus allowing their convenient isolation ( Astle et al. Chem. Biol. 2010 , 17 , 38 - 45 ). However, recent work in our laboratory and others has shown that magnetic hit recovery can result in the isolation of large numbers of false positives and has also suggested that many true hit beads are missed. In this study, we employ a well-defined model system to examine the efficiency of various magnetic hit isolation protocols. We show that the choice of reagents and the particular operations employed are critical for optimal results.
authors.affiliation: Opko Health, Inc. , RF Building, Jupiter, Florida 33458, United States.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Mendes
authors.affiliation: Opko Health, Inc. , RF Building, Jupiter, Florida 33458, United States.
authors.forename: J M
authors.initials: JM
authors.lastname: Ndungu
authors.affiliation: Departments of Chemistry and Cancer Biology, The Scripps Research Institute , 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Lorraine F
authors.initials: LF
authors.lastname: Clark
authors.affiliation: Departments of Chemistry and Cancer Biology, The Scripps Research Institute , 130 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kodadek
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005189:False Positive Reactions; D007074:Immunoglobulin G / Q000737:chemistry; D007202:Indicators and Reagents; D008024:Ligands; D008280:Magnetics; D008863:Microspheres; D008956:Models, Chemical; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis; D011506:Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D054852:Small Molecule Libraries
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D011506:Proteins; D054852:Small Molecule Libraries
keywords: antibody; magnetic recovery; one-bead−one-compound library; peptides; peptoids; screening