Taxonomic identification of pathogenic microorganisms and their toxic proteins

Abstract

The present invention describes a method for the binding of pathogenic microorganisms and their toxic proteins with ligands that have been covalently tethered at some distance from the surface of a substrate: distances of at least fifteen Å are required for microorganism binding ligand tethers and at least six Å are required for protein binding ligand tethers. The ligands described herein include heme compounds, siderophores, polysaccharides, and peptides specific for toxic proteins, outer membrane proteins and conjugated lipids. Non-binding components of the solution to be analyzed are separated from the bound fraction and binding is confirmed by detection of the analyte via microscopy, fluorescence, epifluorescence, luminescence, phosphorescence, radioactivity, or optical absorbance. By patterning numerous ligands in an array on a substrate surface it is possible to taxonomically identify the microorganism by analysis of the binding pattern of the sample to the array.

73 . A method for identification of a protein comprising: 
 (a) exposing the solution containing microorganisms to conditions resulting in the rupture of the microorganism and spilling of the contents into the solution selected from the group comprising chemical treatment, holins, enzymatic treatment, freeze-thaw cycling, bacteriophage infection, and physical treatment.    (b) exposing a solution containing the protein analyte to a ligand specific for the analyte of interest that has been covalently tethered with a photostable linker to a substrate surface;    (c) separating the bound analyte from the non-binding components of the solution containing the analyte by physical separation; and    (d) interrogating the ligand-tethered substrate surface for analyte binding wherein the ligand is tethered at a distance of at least six Å from the substrate surface for the capture of proteins.    
     
     
         74 . The method of  claim 73 , wherein the ligand is a peptide specific for the protein of interest.  
     
     
         75 . The method of  claim 73 , wherein the detection is via the intrinsic fluorescence of the captured protein.