Colloidal compositions for solid phase biomolecular analytical, preparative and identification systems

Abstract

A liquid composition comprising a colloidal suspension of a biomolecule-binding matrix material (preferably nitrocellulose) dispersed in a liquid, with particles of the matrix material being of a defined particle size, and replicate copies of a biomolecule, e.g., protein or nucleic acid probes, which are distributed, preferably uniformly, throughout the colloidal suspension and are bound to the matrix material particles, is disclosed. The liquid composition of the invention can be used directly for sample analysis or preparation of biomolecules, or aliquots of the composition can be spotted onto a support to form a microporous matrix system or microarray for analysis or preparation of biomolecules. Compositions and microarrays according to the invention are useful in any type of analytical or preparative procedure relating to biomolecules. They are particularly useful, e.g., in methods for detecting a biomolecule analyte in a liquid sample, methods for determining the presence of a particular nucleic acid sequence within a liquid sample and methods for determining the presence of a drug candidate molecule in a liquid sample. The invention further comprises kits for practicing the various methods of the invention.

1 - 32 . (canceled) 
   
   
       33 . A microarray having a multiplicity of spots, wherein the composition of each of said spots comprises a biomolecule-binding matrix material, said matrix material being prepared in colloidal form, a first biomolecule bound to said matrix material, and a second biomolecule bound to said matrix material, wherein the concentration of said matrix material and the concentration of said first biomolecule may vary among the said spots, and wherein said spots are comprised of a multiplicity of particles forming a porous structure to enhance contact with and detection of an analyte. 
   
   
       34 . The microarray of  claim 33 , wherein said first and second biomolecules are biomolecular probes. 
   
   
       35 . The microarray of  claim 33 , wherein said first biomolecule is a biomolecule probe and said second biomolecule is a blocking biomolecule, wherein said blocking biomolecule blocks sites on said biomolecule-binding matrix material not occupied by said biomolecule probe. 
   
   
       36 . The microarray of  claim 33 , further comprising in at least one spot a first reference dye, wherein the concentration of said first reference dye has a defined quantitative relationship to the concentration of said biomolecule binding matrix material. 
   
   
       37 . The microarray of  claim 33 , further comprising in at least one spot a second reference dye, wherein the concentration of said second reference dye has a defined quantitative relationship to the concentration of said first biomolecule. 
   
   
       38 . (canceled) 
   
   
       39 . A microarray on a solid support surface, said microarray having a multiplicity of spots, wherein the composition of each spot comprises:
 a biomolecule-binding colloidal matrix material; and   a first biomolecule bound to said matrix material,   wherein the thickness of each of said spots is determined so as to maximize the detection of an analyte, and said spots comprise a multiplicity of particles forming a porous structure to enhance contact with and detection of an analyte.   
   
   
       40 . The microarray of  claim 39 , wherein the thickness of each of said spots is less than 5 μm, less than 2.5 μm or less than 1 μm. 
   
   
       41 . The microarray of  claim 39 , wherein:
 the volume of each of said multiplicity of spots is less than 100 nl.   
   
   
       42 . The microarray of  claim 41 , wherein the volume of each of said spots is less than 50 nl or less than 20 nl. 
   
   
       43 - 54 . (canceled) 
   
   
       55 . The microarray of  claims 33  or  39 , wherein the biomolecules are selected from the group consisting of proteins, peptides and oligonucleotides. 
   
   
       56 . The microarray of  claims 33  or  39 , wherein the particles are substantially spherical in shape. 
   
   
       57 . The microarray of  claim 39  wherein the solid support surface is selected from the group consisting of slides, tubes and microwell plates.