Assay for High-Throughput Identification of Therapeutic Compounds

Abstract

A solid supported branched linker assay system, including an alpha compound and a beta compounds reversibly tethered to a solid support; a branched linker coupled to the solid support that tethers the alpha and beta compounds to the solid support; the branched linker having two cleavable linkers that are chemically distinct from one another, wherein a first chemically distinct linker tethers the β compound to the branched linker and a second chemically distinct linker tethers the α compound to the branched linker; and at least two means for cleaving the chemically distinct linkers, wherein a first cleavage means is configured to selectively cleave a first chemically distinct linker and a second cleavage means is configured to selectively cleave a second chemically distinct linker.

What is claimed: 
     
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         11 . A method of identifying effective therapeutic compounds comprising:
 a.) reversibly coupling an alpha compound and a beta compound to a solid support through a branched linker, wherein the branched linker comprises two cleavable linkers that are chemically distinct from one another;   b) the two cleavable linkers further comprising a first chemically distinct linker that tethers the beta compound to the branched linker and a second chemically distinct linker that tethers the alpha compound to the branched linker; and   b.) providing at least two means for cleaving the chemically distinct linkers, wherein a first cleavage means is configured to selectively cleave the first chemically distinct linker and a second cleavage means is configured to selectively cleave the second chemically distinct linker.   
     
     
         12 . The method of  claim 11 , wherein the solid support is a polyethylene-grafted polystyrene bead. 
     
     
         13 . The method of  claim 11 , wherein the alpha or beta compounds are peptides or antimicrobial peptides. 
     
     
         14 . The method of  claim 11 , wherein the alpha or beta compounds are peptoids. 
     
     
         15 . The method of  claim 11 , wherein the alpha and beta compounds are substantially identical. 
     
     
         16 . The method of  claim 11 , wherein:
 a.) the first chemically distinct linker comprises a disulfide; and   b.) the first cleavage means comprises a reducing reagent.   
     
     
         17 . The method of  claim 11 , wherein:
 a.) the second chemically distinct linker comprises a methionine; and   b.) the second cleavage means comprises cyanogen bromide.   
     
     
         18 . The method of  claim 11 , further comprising screening the therapeutic effectiveness of the beta compound and assessing the identity of the alpha compound, wherein:
 a.) a growth media is inoculated with cells of interest;   b.) adding to the growth media the alpha and beta compounds tethered to the solid support and the first cleavage means to form a growth media complex;   c.) incubating the growth media complex during which the cells grow within and on the growth media and the first chemically distinct linker is cleaved, thereby removing the beta compound from the support structure;   d.) assessing the therapeutic effectiveness of the cleaved compound within the growth media complex by analyzing the degree of cell growth inhibition that surrounds the solid support.   e.) removing the solid support and the remaining alpha compound tethered thereto from the growth media;   f.) adding the second cleavage means to the solid support and the alpha tethered thereto to cleave the second chemically distinct linker and release the alpha compound from the support media;   g.) determining the identity of the alpha compound.   
     
     
         19 . The method of  claim 18 , wherein the cells of interest are microorganisms. 
     
     
         20 . The method of  claim 18 , wherein the growth media is soft agar.