Liquid tissue preparation from histopathological processed biological samples, tissues and cells

Abstract

The current invention provides a method for directly converting histopathologically processed biological samples, tissues, and cells into a multiuse biomolecule lysate. This method allows for simultaneous extraction, isolation, solubilization, and storage of all biomolecules contained within the histopathologically processed biological sample, thereby forming a representative library of said sample. This multi-use biomolecule lysate is dilutable, soluble, capable of being fractionated, and used in any number of subsequent experiments.

1 .- 18 . (canceled) 
     
     
         19 . A method of detecting one or more analytes in a multi-use biomolecule lysate suspected of containing said one or more analytes, comprising the steps of:
 (a) contacting a multi-use biomolecule lysate with an array, wherein said array comprises one or more capture reagents of known binding specificity immobilized on a support surface in a positionally distinguishable manner; and   (b) detecting the binding or absence of binding of one or more analytes in said lysate to said immobilized capture reagents;   wherein the multi-use biomolecule lysate is prepared by   heating a composition comprising a histopathologically processed biological sample and a reaction buffer at a temperature and a time sufficient to negatively affect protein cross-linking in said biological sample, and   treating the resulting composition with an effective amount of a proteolytic enzyme for a time sufficient to disrupt the tissue and cellular structure of said biological sample.   
     
     
         20 . The method according to  claim 19 , wherein at least one of said analytes is a protein. 
     
     
         21 . The method according to  claim 20 , wherein said one or more capture reagents is selected from the group consisting of antibodies and antibody fragments, single domain antibodies, engineered scaffolds, peptides, nucleic acid aptamers, a receptor moiety, affinity reagents, small molecules, and protein ligands. 
     
     
         22 . The method according to  claim 20 , wherein the support surface comprises a material selected from the group consisting of glass, derivitized glass, silicon, derivitized silicon, porous silicon, plastic, a nitrocellulose membranes, a nylon membranes, and a PVDF membranes. 
     
     
         23 . A method of analyzing a plurality of multi-use biomolecule lysates obtained from a plurality of histopathologically processed biological samples, comprising the steps of
 (a) immobilizing a plurality of multi-use biomolecule lysates obtained from a histopathologically processed sample on a support surface, wherein each lysate is immobilized at a discrete location on said surface;   (b) contacting said support surface with a reagent of known binding affinity; and   (c) detecting the presence or absence of binding of said reagent of known binding affinity at said discrete locations on said support surface.   
     
     
         24 . The method according to  claim 23 , wherein the multi-use biomolecule lysate is spotted onto the support surface by a method selected from the group consisting of manual spotting, ink-jetting, robotic contact printing, robotic noncontact printing and piezoelectric spotting. 
     
     
         25 . The method according to  claim 23 , wherein said reagent of known binding affinity is selected from the group consisting of antibodies and antibody fragments, single domain antibodies, engineered scaffolds, peptides, nucleic acid aptamers, a receptor moiety, affinity reagents, small molecules, and protein ligands. 
     
     
         26 . The method according to  claim 23 , wherein said support surface comprises a material selected from the group consisting of glass, derivitized glass, silicon, derivitized silicon, porous silicon, plastic, nitrocellulose membranes, nylon membranes, and PVDF membranes. 
     
     
         27 . The method according to  claim 19  wherein at least one of said analytes is a nucleic acid. 
     
     
         28 . The method according to  claim 27 , wherein said multi-use biomolecule lysate is subjected to a fractionation step prior to contacting said lysate with said array. 
     
     
         29 . The method according to  claim 27 , wherein said nucleic acid comprises RNA. 
     
     
         30 . The method according to  claim 27 , wherein said nucleic acid comprises DNA. 
     
     
         31 . The method according  claim 28 , wherein an RNA-containing fraction of each lysate is immobilized on said support surface. 
     
     
         32 . The method according to  claim 19 , wherein the detecting step (b) is carried out using a detection reagent that specifically binds to one or more of the analytes suspected to be present in said sample. 
     
     
         33 . The method according to  claim 32 , wherein said detection reagents are proteins. 
     
     
         34 . The method according to  claim 23 , wherein one or more of said multi-use biomolecule lysates is subjected to a fractionation step prior to immobilizing one or more of the resulting fractions on said surface. 
     
     
         35 . The method according to  claim 34 , wherein the a nucleic acid fraction of one or more lysates is immobilized on said surface. 
     
     
         36 . The method according to  claim 35 , wherein said nucleic acid fraction is an RNA-containing fraction. 
     
     
         37 . The method according to  claim 35 , wherein said nucleic acid fraction is a DNA-containing fraction. 
     
     
         38 . The method according to  claim 34 , wherein said fraction is a protein-containing fraction. 
     
     
         39 . (canceled)