Stable, water-soluble quantum dot, method of preparation and conjugates thereof

Abstract

A method for manufacturing powdered quantum dots comprising the steps of: a) reacting quantum dots comprising a core, a cap and a first ligand associated with the outer surfaces thereof with a second ligand, the second ligand displacing the first ligand and attaching to the outer surfaces of the quantum dots, b) isolating the quantum dots having the attached second ligand from the reaction mixture, c) reacting the isolated quantum dots having the attached second ligand with a small organic molecule whereby the small organic molecule attaches to the second ligand, d) reacting the quantum dots having the attached small organic molecule with a cross-linking agent to cross-link the small organic molecule attached to the second ligand with an adjacent second ligand attached to the surfaces of the quantum dots, e) isolating the quantum dots formed in step (d); and f) drying the isolated quantum dots to form powdered quantum dots. The invention includes the quantum dots.

1 . A method for manufacturing powdered quantum dots comprising the steps of: 
 a) reacting quantum dots comprising a core, a cap and a first ligand associated with the outer surfaces thereof with a second ligand, the second ligand displacing the first ligand and attaching to the outer surfaces of the quantum dots,    b) isolating the quantum dots having the attached second ligand from the reaction mixture,    c) reacting the isolated quantum dots having the attached second ligand with a small organic molecule whereby the small organic molecule attaches to the second ligand,    d) reacting the quantum dots having the attached small organic molecule with a cross-linking agent to cross-link the small organic molecule attached to the second ligand with an adjacent second ligand attached to the surfaces of the quantum dots,    e) isolating the quantum dots formed in step (d); and    f) drying the isolated quantum dots to form stable, oxidation-resistant, powdered quantum dots.    
     
     
         2 . The method of  claim 1 , wherein the second ligand is a compound having a formula ROC—(CH 2 ) n —COOH, where R is a thiol, and n is a whole integer in the range of 8 to 13.  
     
     
         3 . The method of  claim 1 , wherein the second ligand comprises a hydrophilic moiety.  
     
     
         4 . The method of  claim 3 , wherein the second ligand is mercaptoundecanoic acid.  
     
     
         5 . The method of  claim 1 , wherein the small organic molecule is selected from molecules containing at least two amino groups and one carboxylic acid group.  
     
     
         6 . The method of  claim 5 , wherein the organic molecule is an amino acid.  
     
     
         7 . The method of  claim 6 , wherein the amino acid is lysine.  
     
     
         8 . The method of  claim 5 , wherein the small organic molecule is a compound having a formula R 1 OC—(CH 2 ) n COR 2  where each of R 1  and R 2  is an amine group and n is a whole integer in the range of 8 to 13.  
     
     
         9 . The method of  claim 1 , wherein the second ligand is supplied in a molar excess of at least about 4,000 compared to the quantum dot having an attached first ligand population.  
     
     
         10 . The method of  claim 1  further comprising the steps of: 
 a) dissolving the powdered quantum dots in an aqueous solution; and    b) contacting the dissolved quantum dots with a biomolecule whereby the quantum dot and the biomolecule form a conjugate.    
     
     
         11 . The method of  claim 10 , wherein the biomolecule is selected from the group comprising: a protein or an antigenically reactive fragment thereof, an antibody or an antigenically reactive fragment thereof and a nucleic acid.  
     
     
         12 . A method of detecting a biomolecule in a sample comprising the steps of: 
 a) contacting the sample with the conjugate prepared by the method of  claim 10 , wherein the biomolecule to be detected specifically binds to the biomolecule conjugated to the quantum dots; and    b) detecting luminescence, wherein the detection of luminescence is indicative of the presence of the biomolecule in the sample.    
     
     
         13 . The method of  claim 11 , wherein the biomolecule to be detected is selected from the group comprising: a protein, an antigenically reactive fragment of the protein, an antibody, an antigenically reactive fragment of the antibody and a nucleic acid.  
     
     
         14 . A water-soluble powder comprising quantum dots manufactured according to the method of  claim 1 .  
     
     
         15 . The method of  claim 2  further comprising the steps of: 
 a) dissolving the powdered quantum dots in an aqueous solution; and    b) contacting the dissolved quantum dots with a biomolecule whereby the quantum dot and the biomolecule form a conjugate.    
     
     
         16 . The method of  claim 15 , wherein the biomolecule is selected from the group comprising a protein or an antigenically reactive fragment thereof, an antibody or an antigenically reactive fragment thereof and a nucleic acid.  
     
     
         17 . A method of detecting a biomolecule in a sample comprising the steps of: 
 a) contacting the sample with a conjugate prepared by the method of  claim 15 , whereby the biomolecule to be detected specifically binds to the biomolecule conjugated to the quantum dot; and    b) detecting luminescence, wherein the detection of luminescence is indicative of the presence of the biomolecule in the sample.    
     
     
         18 . A quantum dot comprising: 
 a core,    a cap,    a ligand attached to the outer surface of the cap, said ligand being a compound having a formula ROC—(CH 2 ), —COOH, where R is an organic molecule with a hydrophilic moiety and n is a whole integer in the range of 8 to 13; and,    a cross-linker comprising a molecule having a formula R 1 OC—(CH 2 ) n —COR 2  where each of R 1  and R 2  is an amine group and n is a whole integer in the range of 8 to 13, said cross-linker joining two adjacent ligands.    
     
     
         19 . A quantum dot of  claim 18  further comprising a biomolecule conjugated to an exposed polar group on the cross-linker.  
     
     
         20 . The quantum dot of  claim 18  that remains monodispersed in an aqueous solution for at least 10 days.