High-throughput receptor-binding methods for somatostatin receptor 2.
Three high-throughput screening methods for quantitating 125I-SS14 binding to human somatostatin receptor 2 (hSST2) have been developed. Microplate-based separation assays were performed in Packard Unifilter and Millipore Multiscreen plates. A homogeneous ligand-binding assay was developed by employing wheat germ agglutinin (WGA)-coated Flashplates. Apparent dissociation constants for 125I-SS14 binding to hSST2 were obtained with each method. IC(50) values were determined for 12 compounds using each of the methods. Similar IC(50) values were obtained for each compound with all of the methods. The WGA-Flashplate is suitable for fully automated high-throughput screening whereas the Unifilter and Multiscreen methods are more suitable for semiautomated and manual screening applications.
Department of Atherosclerosis and Endocrinology, Merck Research Laboratories, Rahway, NJ 07065, USA. elizabeth_birzin@merck.com
Elizabeth T
ET
Birzin
Susan P
SP
Rohrer
D000818:Animals; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology; D001667:Binding, Competitive; D016466:CHO Cells; D006224:Cricetinae; D004353:Drug Evaluation, Preclinical; D006801:Humans; D020128:Inhibitory Concentration 50; D008024:Ligands; D008851:Micropore Filters; D015282:Octreotide / Q000494:pharmacology; D011485:Protein Binding; D011498:Protein Precursors / Q000494:pharmacology; D011869:Radioligand Assay; D017481:Receptors, Somatostatin / Q000378:metabolism*; D013004:Somatostatin / Q000494:pharmacology; D053561:Somatostatin-28; D014909:Wheat Germ Agglutinins / Q000737:chemistry / Q000378:metabolism
D018931:Antineoplastic Agents, Hormonal; D008024:Ligands; D011498:Protein Precursors; D017481:Receptors, Somatostatin; D014909:Wheat Germ Agglutinins; D013004:Somatostatin; D053561:Somatostatin-28; C093733:somatostatin receptor 2; D015282:Octreotide