Encoded chemical synthesis coupled to screening: "Pot Assay".
A variety of screening methodologies is available to identify lead compounds. Screening methods that would permit the direct use of libraries made via the Radiofrequency Encoded Combinatorial chemistry paradigm (each individual small molecule in the library is presented separately on an individual encoded support) have the potential to diminish burdensome steps in this process. Here we report on our studies leading to such a direct method, which we have termed a Pot Assay. Pot Assay is a multiplex assay, which simultaneously measures specific binding of a number of ligands to at least one target. Pot Assay uses specific radiofrequency signals to decode compounds that are high affinity binders. We validated this approach by evaluating the interaction of biotin and its analogs with labeled streptavidin. This report introduces Pot Assay as a rapid, simple, sensitive and accurate format for identifying active members of libraries synthesized on solid supports. The success of this study demonstrates the power of coupling Radiofrequency Encoded Combinatorial chemistry and screening. This assay format may be applied to a wide range of screens that are based on binding events: ligand/receptor, inhibitor/enzyme, antigen/antibody, protein/protein, DNA/protein, and RNA/DNA.
Parandoosh
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IRORI, La Jolla, CA 92037-1031, USA. parandoosh@aol.com
Knowles
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Xiao
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Zhao
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C
David
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Nova
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MP
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D001710:Biotin / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D002626:Chemistry, Pharmaceutical / Q000295:instrumentation / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation* / Q000379:methods*; D007457:Iodine Radioisotopes; D008163:Luminescent Measurements; D011846:Radio Waves; D019809:Streptavidin / Q000378:metabolism*
D007457:Iodine Radioisotopes; D001710:Biotin; C004749:desthiobiotin; D019809:Streptavidin; D000469:Alkaline Phosphatase