Supramolecular bioconjugates

Abstract

The invention relates to supramolecular bioconjugates and to methods for assembling and utilizing supramolecular bioconjugates. Supramolecular bioconjugates comprise a plurality of first nucleic acids and a plurality of mediators wherein each mediator comprises a second nucleic acid complementary to a sequence within said plurality of first nucleic acids. To assemble a supramolecular bioconjugate, one or more sets of bioreactive agents are coupled to the plurality of mediators, forming a plurality of bioreactive complexes. The plurality of bioreactive complexes are hybridized to the plurality of first nucleic acids to form the supramolecular bioconjugate. Bioconjugates can be used to detect and isolate targets, to screen samples for targets such as antigens, to treat patients with multiple agents or to diagnose disorders in the form of a kit.

1 . A method for assembling a supramolecular bioconjugate comprising the steps of:
 a) providing a first bioreactive complex said bioreactive complex comprising a first nucleic acid, said first nucleic acid attached to a first bioreactive agent, said first bioreactive agent comprising a first binding agent selected from the group consisting of protein A, an antibody, an Fc fragment, a Fab fragment, an antigen, a nucleic acid binding protein, and a cell adhesion molecule, wherein said first bioreactive agent is coupled to a molecule binding to said first bioreactive agent;   b) providing at least a second bioreactive complex comprising at least two second nucleic acids, wherein one of the at least two second nucleic acids serves as a first mediator nucleic acid that comprises a nucleic acid sequence that is complementary to a sequence within said first nucleic acid, and wherein said at least two second nucleic acids are each attached to a second bioreactive agent, said second bioreactive agent comprising a second binding agent selected from the group consisting of protein A, an antibody, an Fc fragment, a Fab fragment, an antigen, a nucleic acid binding protein, and a cell adhesion molecule; and   c) hybridizing in one step said complementary nucleic acids of said second bioreactive complex to said first nucleic acid of said first bioreactive complex to form a supramolecular bioconjugate, wherein at least one of the second bioreactive agent is selected to be able to react primarily with a desired bioreactive target, while part of the supramolecular bioconjugate.   
     
     
         2 . The method of  claim 1  wherein the first and the second nucleic acid is between about 10 to about 1000 nucleotides in length. 
     
     
         3 . The method of  claim 1 , wherein the first nucleic acid is between about 20 to about 60 nucleotides in length and the one or more second nucleic acids are between about 20 to about 60 nucleotides in length. 
     
     
         4 . The method of  claim 1 , wherein the first and the second nucleic acid is between 10 to 600 nucleotides in length. 
     
     
         5 . The method of  claim 1 , wherein all the first nucleic acids contain a common nucleic acid sequence. 
     
     
         6 . The method of  claim 1 , wherein all the second nucleic acids contain a common nucleic acid sequence. 
     
     
         7 . The method of  claim 1 , wherein the first nucleic acid and the second nucleic acid is DNA, RNA or PNA. 
     
     
         8 . The method of  claim 1 , wherein the first and the second bioreactive agent is selected from the group consisting of antibodies, antigens, lectins, cytokines, nucleic acids, aptamers, metals, metabolites, metabolic analogs, enzymes, cofactors, bacteriostatic agents, bacteriocidal agents, amino acids, peptides, proteins, chemotherapeutic agents, avidin, streptavidin, biotin, toxins, cytotoxic agents, substrates, antiviral agents, antifungal agents, monosaccharides, polysaccharides or combinations or fragments thereof. 
     
     
         9 . A method for forming a nucleic acid-directed array of immobilized supramolecular bioconjugates comprising the steps of:
 a) attaching a first nucleic acid to a support in a regiospecific manner;   b) providing at least a second nucleic acid complementary to a sequence within said first nucleic acid, wherein said second nucleic acid is coupled to a bioreactive agent, wherein said second bioreactive agent is a non-nucleic acid bioreactive agent and contains a separate binding moiety that is selected to further react with a specific target;   c) hybridizing in one step said first nucleic acid to said second nucleic acid to form a supramolecular bioconjugate; and   d) repeating steps (a)-(d) to form a plurality of immobilized supramolecular bioconjugates, wherein the separate binding moiety of each of said bioreactive agent is selected to further react with a specific target.   
     
     
         10 . The method of  claim 9 , wherein the second and the third bioreactive agent further comprises a binding agent. 
     
     
         11 . The method of  claim 10 , wherein the binding agent is selected from the group consisting of avidin, streptavidin, and biotin. 
     
     
         12 . The method of  claim 9 , wherein the support is a ceramic, a glass, a plastic, a metal, a resin, a gel, an elastic, a wood or a combination thereof. 
     
     
         13 . The method of  claim 9 , wherein the second nucleic acid is between 10 to 600 nucleotides in length. 
     
     
         14 . The method of  claim 9 , wherein the support is three dimensional. 
     
     
         15 . A supramolecular bioconjugate comprising;
 a) a first bioreactive complex said bioreactive complex comprising a first nucleic acid, said first nucleic acid attached to a first bioreactive agent, said first bioreactive agent comprising a first binding agent selected from the group consisting of protein A, an antibody, an Fc fragment, a Fab fragment, an antigen, a nucleic acid binding protein, and a cell adhesion molecule, wherein said first bioreactive agent is coupled to a molecule binding to said first bioreactive agent; and   b) at least a second bioreactive complex comprising at least two second nucleic acids, wherein one of the at least second nucleic acids serves as a first mediator nucleic acid that comprises a nucleic acid sequence that is complementary to a sequence within said first nucleic acid, said at least two second nucleic acids attached to a second bioreactive agent, said second bioreactive agent comprising a second binding agent selected from the group consisting of protein A, an antibody, an Fc fragment, a Fab fragment, an antigen, a nucleic acid binding protein, and a cell adhesion molecule, and wherein the first and the second nucleic acids are at least partially hybridized with each other.   
     
     
         16 . The supramolecular bioconjugate of  claim 15 , wherein the supramolecular bioconjugate has been treated with an agent which degrades at least a portion of the first and/or the second nucleic acid. 
     
     
         17 . The supramolecular bioconjugate of  claim 15 , further comprising at least a third bioreactive complex comprising a third nucleic acid attached to a third bioreactive agent, wherein the third nucleic acid serves as a second mediator nucleic acid that comprises a nucleic acid sequence that is at least partially complementary to the second or the first nucleic acid, and wherein the third bioreactive agent comprises a third binding agent selected from the group consisting of protein A, an antibody, an Fc fragment, a Fab fragment, an antigen, a nucleic acid binding protein, and a cell adhesion molecule. 
     
     
         18 . The supramolecular bioconjugate of  claim 15 , wherein each of the first and the second nucleic acids is between about 20 to about 600 nucleotides in length. 
     
     
         19 . The supramolecular bioconjugate of  claim 15 , which has been treated with an agent which degrades nucleic acid. 
     
     
         20 . The supramolecular bioconjugate of  claim 19  wherein the agent is an acid, a base or a nuclease. 
     
     
         21 . The supramolecular bioconjugate of  claim 15  wherein the first and the second bioreactive agent is selected from the group consisting of antibodies, antigens, lectins, cytokines, nucleic acids, aptamers, metals, metabolites, metabolic analogs, enzymes, cofactors, bacteriostatic agents, bacteriocidal agents, amino acids, peptides, proteins, chemotherapeutic agents, avidin, streptavidin, biotin, toxins, cytotoxic agents, substrates, antiviral agents, antifungal agents, monosaccharides, polysaccharides or combinations or fragments thereof. 
     
     
         22 . The supramolecular bioconjugate of  claim 15  further comprising a solid support. 
     
     
         23 . The supramolecular bioconjugate of  claim 15  further comprising a pharmaceutically active agent and a pharmaceutically acceptable carrier. 
     
     
         24 . The supramolecular bioconjugate of  claim 22 , wherein the solid support comprises at least 10 immobilized supramolecular bioconjugates. 
     
     
         25 . The supramolecular bioconjugate of  claim 22 , wherein the solid support comprises at least 1,000 immobilized supramolecular bioconjugates. 
     
     
         26 . The supramolecular bioconjugate of  claim 22 , wherein the solid support comprises at least 100,000 immobilized supramolecular bioconjugates. 
     
     
         27 . The supramolecular bioconjugate of  claim 22 , wherein the solid support comprises at least 1,000,000 immobilized supramolecular bioconjugates.