Material for supported synthesis and method for growing oligonucleotides or peptides

Patent metadata

Field	Value
Publication number	US-9359396-B2
Application number	US-201214131490-A
Country	US
Kind code	B2
Filing date	Jul 10, 2012
Priority date	Jul 12, 2011
Publication date	Jun 7, 2016
Grant date	Jun 7, 2016

Abstract

The invention concerns a material composed of a porous support on which functionalized nanoparticles are grafted by covalent bonding, characterized in that at least part of the nanoparticles grafted by covalent bonding is housed inside surface pores of the support, and in that the support is silica-based and is in the form of porous particles of heterogeneous shape and size, the size of the particles being larger than 1 μm and preferably within the range of 5 to 200 μm.
The invention further concerns a growth method for oligonucleotides or peptides characterized in that growth is performed on a material formed of a porous support on which functionalized nanoparticles are grafted by covalent bonding, characterized in that at least a part of the nanoparticles grafted by covalent bonding is housed inside surface pores of the support.

First claim

The invention claimed is:

1. A material composed of a porous support on which functionalized nanoparticles are grafted by covalent bonding, characterized in that at least a portion of the nanoparticles grafted by covalent bonding is housed inside surface pores of the support, and in that the support is silica-based and is in the form of porous particles of heterogeneous shape and size, the size of the particles being larger than 1 μm.

2. The material according to claim 1 , characterized in that the support is a mineral glass containing silica or a silicate.

3. The material according to claim 1 , characterized in that the support is composed exclusively of silica or is composed exclusively of a silicate in which the silica is in a mixture with B 2 O 3 to form borosilicates or Al 2 O 3 to form aluminosilicates, optionally in a mixture with Na 2 O.

4. The material according to claim 1 , characterized in that the minimum mean size of the surface openings formed by the surface pores of the support is at least 3 times equal to the size of the nanoparticles.

5. The material according to claim 1 , characterized in that the mean diameter of the pores of the support is in the range of 30 to 600 nm.

6. The material according to claim 5 , characterized in that the mean diameter of the pores of the support is in the range of 100 to 400 nm.

7. The material according to claim 1 , characterized in that the support has homogeneous porosity corresponding to the fact that at least 80% of the pores have a diameter whose value does not vary by more than 10% compared with the mean diameter of the pores.

8. The material according to claim 1 , characterized in that the support has a volume of pores per gram of 0.5 to 1.5 mL/g.

9. The material according to claim 1 , characterized in that the support has a specific surface area in the range of 5 to 80 m 2 /g.

10. The material according to claim 1 , characterized in that the nanoparticles have a mean size in the range of 2 to 100 nm.

11. The material according to claim 1 , characterized in that at least 80% of the grafted nanoparticles are directly bonded by a covalent bond to the support without any intermediate nanoparticles ensuring the said bonding.

12. The material according to claim 11 , characterized in that at least 90% of the grafted nanoparticles are directly bonded by a covalent bond to the support.

13. The material according to claim 1 , characterized in that the nanoparticles are nanoparticles of silica, gold, silver, graphite, fluorescent nanocrystals, polymer nanoparticles of polystyrene, polypropylene or polybutadiene type, or metal oxide nanoparticles, optionally coated to enable the functionalization thereof.

14. The material according to claim 13 , characterized in that the nanoparticles are nanoparticles of tin oxide, gadolinium oxide, aluminium oxide, titanium oxide, cerium oxide, niobium oxide, erbium oxide, ytterbium oxide, terbium oxide, dysprosium oxide or europium oxide, or of a mixture of these oxides, optionally coated to enable the functionalization thereof.

15. The material according to claim 1 , characterized in that the nanoparticles are functionalized by reactive functions chosen from among alcohols, amines and thiols allowing the growth of oligonucleotides or peptides.

16. The material according to claim 1 , characterized in that the nanoparticles are grafted on to the support by a cleavable bond cleavable under conditions not deteriorating the biological ligands.

17. The material according to claim 16 , wherein the cleavable bond is selected from the group consisting of an ester bond, alkoxysilyl bond, disulphide bridge, 4-(2-hydroxyethyl)-3-nitrobenzoic acid bond, N-[9-(hydroxymethyl)-2-fluorenyl]succinamic acid bond, thiophosphoramidatehydroxypropyl bond and an amide bond activated by a 2-propane diol group.

18. The material according to claim 1 , characterized in that the covalent bond between the nanoparticles and the support is obtained by coupling between an arm with an acid terminal function positioned on the support, chosen among: —(CH2)n-NH—CO—(CH2)m-COON, —(CH2)n-COOH, —(CH2)n-NH—(CH2)m-COOH, with n being an integer ranging from 2 to 18 and m an integer ranging from 2 to 6, and
an arm with an alcohol terminal function positioned on the nanoparticles, chosen among:
—(CH2)q-NH—CO—NH—(CH2)r-OH, —(CH2)s-OH, —(CH2)q-O—[(CH2)2-O]p-Het-CH2)q-CONH—(CH2)r-OH,—(CH2)q-NH—(CH2)r-OH, —(CH2)q-CHOH—CH2-NH—(CH2)p-OH, —(CH2)q-[CH—(CH2OH)]—NH—(CH2)p-OH, —(CH2)q-CHOH—CH2-O—(CH2)p-OH, —(CH2)q-[CH—(CH2OH)]—O—(CH2)p-OH, —(CH2)q-CHOH—CH2-O—CO—NH—(CH2)p-OH
with s being an integer ranging from 2 to 18, p an integer ranging from 1 to 6, q an integer ranging from 1 to 12, and r an integer ranging from 1 to 12.

19. The material according to claim 1 , characterized in that the nanoparticles are functionalized with free hydrophilic organic molecules or with fluorescent organic molecules.

20. The material according to claim 19 , characterized in that the nanoparticles are functionalized with free hydrophilic organic molecules chosen among polyethylene glycol and polyethylamine, or with fluorescent organic molecules chosen among fluorescein, rhodamine and cyanines.

21. The material according to claim 1 , characterized in that the nanoparticles are functionalized, by covalent bonding, with biological ligands chosen among peptides and oligonucleotides.

22. The material according to claim 21 , characterized in that it has a high density of biological ligands grafted on the surface of the nanoparticles, corresponding to more than 0.1 molecule of biological ligand per nm 2 of nanoparticles.

23. The material according to claim 21 , characterized in that the nanoparticles are dissymmetric, so that at least 90% of the biological ligands are grafted on a continuous region representing less than 70% of the surface of the nanoparticle on which they are grafted.

24. The material according to claim 1 , characterized in that the covalent bond between the nanoparticles and the biological ligands corresponds to a peptide or phosphate diester bond.

25. The material according to claim 1 , characterized in that the size of the particles is within the range of 5 to 200 μm.

26. A growth method for oligonucleotides or peptides, characterized in that growth is performed on a material according to claim 1 .

27. The method according to claim 26 , characterized in that growth is performed in an automated synthesizer in which the support is confined in a cell by means of a filter through which flows of argon and reagents circulate alternately.

28. The method according to claim 27 , characterized in that once growth is completed the covalent bond between the support and the nanoparticles is cleaved.

Assignees

• Chaix Carole
• De Crozals Gabriel
• Farre Carole
• Univ Claude Bernard Lyon
• Centre Nat Rech Scient

Inventors

• Chaix Carole
• De Crozals Gabriel
• Farre Carole

Classifications

• B05D7/22

  to internal surfaces, e.g. of tubes · CPC title

• C07K1/042 Primary

  characterised by the nature of the carrier · CPC title

• B01J2219/00632

  Introduction of reactive groups to the surface · CPC title

• B82Y30/00

  Nanotechnology for materials or surface science, e.g. nanocomposites · CPC title

• Y10T428/2982

  Particulate matter [e.g., sphere, flake, etc.] · CPC title