Target molecule attachment to surfaces

Abstract

Method and reagent composition for covalent attachment of target molecules, such as nucleic acids, onto the surface of a substrate. The reagent composition includes groups capable of covalently binding to the target molecule. Optionally, the composition can contain photoreactive groups for use in attaching the reagent composition to the surface. The reagent composition can be used to provide activated slides for use in preparing microarrays of nucleic acids.

1 . A method comprising the steps of: 
 a. providing a support;    b. disposing a reagent composition on the support, wherein the reagent composition comprises a polymeric backbone having more than one thermochemically amine-reactive or sulfhydryl-reactive groups attached thereto, and wherein the reagent composition is configured and arranged to form covalent bonds with functional groups on a target molecule without use of attracting groups to attract the target molecule to the reagent composition;    c. coupling the reagent composition to the support surface to form a reagent composition-coupled support surface; and    d. disposing at least one target molecule on the reagent composition-coupled support surface, wherein the functional groups of the target molecule forms covalent bonds with the reagent composition.    
     
     
         2 . The method according to  claim 1  wherein the target molecule is a biomolecule.  
     
     
         3 . The method according to  claim 2  wherein the biomolecule is a nucleic acid.  
     
     
         4 . The method according to  claim 3  wherein the nucleic acid comprises one or more functional groups selected from the group consisting of amine and sulfhydryl groups.  
     
     
         5 . The method according to  claim 1  wherein the reagent composition comprises one or more photoreactive groups.  
     
     
         6 . The method according to  claim 5  wherein the photoreactive groups are photoreactive aryl ketones selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and heterocyclic analogs of anthrone.  
     
     
         7 . The method according to  claim 5  wherein the thermochemically reactive groups and photoreactive groups are pendent from the polymeric backbone of the reagent composition.  
     
     
         8 . The method according to  claim 1  wherein the polymeric backbone of the reagent composition is selected from the group consisting of acrylics, vinyls, nylons, polyurethanes, and polyethers.  
     
     
         9 . The method according to  claim 1  wherein the thermochemically reactive groups are selected from the group consisting of activated esters, epoxides, azlactones, activated hydroxyls, and maleimide.  
     
     
         10 . The method according to  claim 1  wherein the support is selected from the group consisting of plastic, silicon hydride, or organosilane-pretreated glass or silicone slides.  
     
     
         11 . The method according to  claim 1  wherein the support comprises crystalline thermoplastics or amorphous thermoplastics.  
     
     
         12 . The method according to  claim 1  wherein the step of disposing at least one target molecule is performed with a printing apparatus.  
     
     
         13 . The method according to  claim 1  wherein the step of disposing at least one target molecule comprises disposing a plurality of different target molecules on the reagent composition-coupled support surface.  
     
     
         14 . The method according to  claim 13  performed to fabricate a microarray.  
     
     
         15 . A method according to  claim 14  wherein the microarray includes regions of disposed target molecules which are generally circular in shape, have a diameter of between about 10 microns and about 500 microns, and are separated from other regions in the array by center to center spacing of about 20 microns to about 1000 microns.  
     
     
         16 . The method according to  claim 1  where, in the step of disposing at least one target molecule, the target molecule is disposed on the reagent composition-coupled support surface in a sample having a volume of twenty nanoliters or less.  
     
     
         17 . The method according to  claim 1  where, in the step of disposing at least one target molecule, the target molecule is disposed on the reagent composition-coupled support surface in a quantity in the range of 0.1 femtomoles to 10 nanomoles.