Method for immobilising and/or crystallising biological macromolecules on carbon nanotubes and uses

Abstract

Method for the attachment and/or crystallization of macromolecules, chemical reagents used in the said method, products obtained as well as applications of the said products in the field of materials and of structural biology, in particular as biosensors or as biomaterials. The said method comprises essentially the incubation, without stirring, for at least 15 minutes, of a biological macromolecule in solution with nanotubes of carbon closed at their ends, under suitable temperature and pH conditions.

1 . Method for the attachment and self-organization of biological macromolecules, characterized in that it comprises the incubation, without stirring, for at least 15 minutes, of a biological macromolecule in solution with nanotubes of carbon closed at their ends, under suitable temperature and pH conditions.  
     
     
         2 . Method according to  claim 1 , characterized in that the said biological macromolecules are in particular soluble, membrane or transmembrane proteins, enzymes, antibodies, antibody fragments or nucleic acids.  
     
     
         3 . Method according to  claim 1  or  claim 2 , characterized in that the aid nanotubes of carbon are functionalized by physical adsorption, at their surface, of a chemical reagent of general formula H—E—L,  
       in which: 
 H represents a hydrophilic group selected from the positively or negatively charged groups; ligands or analogues of biological macromolecules; organometallic complexes interacting with amino acids or nucleic acids and whose ligands are optionally functionalized with alkyl groups for bonding to E;  
 E represents a spacer arm, selected from C 1 -C 10  carbon chains, optionally substituted with alkyl groups, having unsaturations or otherwise or polyoxyethylene units which may have or otherwise in the middle of the chain phosphate groups, such as:  
                     
 in which: 
 m represents an integer from 1 to 10,  
 X represents O, NHCO, OCO, COO, CONH, S, CH 2  or NH and constitutes, at the ends of the said carbon chains, organic functions for adhesion of the ester, amide, ether or thioether type;  
 
 L represents a lipid unit with one or more chains of variable length, in the form of C 12 -C 20  having unsaturations or otherwise; an aromatic group of formula Ar 1  or of formula Ar 2 :  
                     
 in which: 
 A represents a hydrogen atom, one of the following groups: alkyl, CF 3 , NO 2 , NH 2 , OH, O-alkyl, S-alkyl, COOH, halogen, an aromatic ring or an aromatic heterocycle in the form of C 4 -C 6 , optionally polysubstituted with electron-donating groups of the alkyl type or electron-attracting groups of the CF 3  or halide type; and  
 Y represents a bond with E.  
 
 
     
     
         4 . Method according to any one of  claims 1  to  3 , characterized in that the said solution consists of a solvent for solubilizing the said biological macromolecules, which is aqueous or aqueous-alcoholic and which optionally contains at least one detergent.  
     
     
         5 . Method according to any one of  claims 1  to  4 , characterized in that the incubation conditions are preferably the following: incubation at room temperature, for 15 minutes to 48 hours, at a pH of between 5.5 and 8.5.  
     
     
         6 . Bionanomaterials, characterized in that they consist of nanotubes of carbon, on which biological macromolecules are self-organized in a crystalline form.  
     
     
         7 . Bionanomaterials according to  claim 6 , characterized in that they are obtained with the aid of a method according to any one of  claims 1  to  5 .  
     
     
         8 . Use of the bionanomaterials according to either of claims  6  and  7 , for the structural study of the biological macromolecules which are associated with them.  
     
     
         9 . Use of the bionanomaterials according to either of claims  6  and  7 , as biological reagent.  
     
     
         10 . Use of the bionanomaterials according to either of claims  6  and  7 , as biosensors or bioconductors.  
     
     
         11 . Chemical reagent capable of being physically adsorbed on nanotubes of carbon, characterized in that it has the general formula H—E—L, in which: 
 H represents a hydrophilic group selected from the positively or negatively charged groups; ligands or analogues of biological macromolecules; organometallic complexes interacting with amino acids or nucleic acids and whose ligands are optionally functionalized with alkyl groups for bonding to E;  
 E represents a spacer arm, selected from C 1 -C 10  carbon chains, optionally substituted with alkyl groups, having unsaturations or otherwise or polyoxyethylene units which may have or otherwise in the middle of the chain phosphate groups, such as:  
                     
 in which: 
 m represents an integer from 1 to 10,  
 X represents O, NHCO, OCO, COO, CONH, S, CH 2  or NH and constitutes, at the ends of the said carbon chains, organic functions for adhesion of the ester, amide, ether or thioether type;  
 
 L represents an aromatic group of formula Ar 1  or of formula Ar 2 :  
                     
 in which: 
 A represents a hydrogen atom, one of the following groups: alkyl, CF 3 , NO 2 , NH 2 , OH, O-alkyl, S-alkyl, COOH, halogen, an aromatic ring or an aromatic heterocycle in the form of C 4 -C 6 , the said rings being optionally polysubstituted with electron-donating groups of the alkyl type or electron-attracting groups of the CF 3  or halide type; and  
 Y represents a bond with E.  
 
 
     
     
         12 . Chemical reagent according to  claim 11 , characterized in that it has one of the following structures:  
       
         
           
           
               
               
           
         
       
     
     
         13 . Chemical reagent according to  claim 11 , characterized in that H is selected from the following organometallic complexes.  
       
         
           
           
               
               
           
         
       
       with R 1 =organic group for bonding to E.