Functionalized surfaces and preparation thereof

Patent metadata

Field	Value
Publication number	US-9895673-B2
Application number	US-201615365826-A
Country	US
Kind code	B2
Filing date	Nov 30, 2016
Priority date	Dec 1, 2015
Publication date	Feb 20, 2018
Grant date	Feb 20, 2018

Abstract

Compositions, devices, methods and systems are provided for differential functionalization of a surface of a structure to support biopolymer synthesis. Provided herein are processes which include use of lamps, lasers, and/or microcontact printing to add functional groups to surfaces for the efficient and uniform synthesis of oligonucleic acids.

First claim

What is claimed is:

1. A method for surface patterning, the method comprising:
applying a first set of molecules to a surface of a structure, wherein each of the first set of molecules binds to the surface and lacks a reactive group capable of binding to a nucleo side;
applying electromagnetic radiation (EMR) to predetermined regions of the surface, wherein the EMR comprises a wavelength from about 100 nm to about 300 nm, wherein application of the EMR results in removal of the first set of molecules at the predetermined regions, thereby defining different loci for oligonucleotide extension; and
synthesizing a plurality of oligonucleotides, wherein each oligonucleotide extends from a different locus, and wherein the different loci are at least about 75% uniform when measured by calculating amplitude of signal variation for oligonucleotides extending from each locus divided by total signal intensity following white light illumination using an optical microscope.

2. The method of claim 1 , wherein greater than about 90% of the first set of molecules are removed at the predetermined regions of the surface following application of EMR.

3. The method of claim 1 , wherein about 100% of the first set of molecules are removed at the predetermined regions of the surface following application of EMR.

4. The method of claim 1 , wherein the predetermined regions have a width of about 1 um to about 500 um.

5. The method of claim 1 , wherein the first set of molecules comprises a fluorosilane.

6. The method of claim 1 , wherein the first set of molecules comprises perfluorooctyltrichlorosilane, (tridecafluoro-1,1,2,2-tetrahydrooctyl)trichlorosilane, or tridecafluoro-1,1,2,2-tetrahydrooctyl)trimethoxysilane.

7. The method of claim 1 , further comprising applying a second set of molecules to the surface after application of the EMR, wherein each of the second set of molecules binds to the predetermined regions of the surface and comprises the reactive group capable of binding to a nucleoside.

8. The method of claim 7 , wherein the second set of molecules comprises an aminosilane.

9. The method of claim 7 , wherein the second set of molecules comprises N-(3-triethoxysilylpropyl)-4-hydroxybutyramide (HAPS), 11-acetoxyundecyltriethoxysilane, n-decyltriethoxysilane, (3-aminopropyl)trimethoxysilane, (3-aminopropyl)triethoxysilane, 3-glycidoxypropyltrimethoxysilane (GOPS), or 3-iodo-propyltrimethoxysilane.

10. The method of claim 1 , wherein each of the oligonucleotides comprises about 25 bases to about 2 kb in length.

11. The method of claim 1 , wherein the oligonucleotides extending from each locus are about 80% uniform when measured by calculating amplitude of signal variation for oligonucleotides extending from each locus divided by total signal intensity following white light illumination using an optical microscope.

12. The method of claim 1 , wherein the EMR comprises a wavelength from about 150 nm to about 200 nm.

13. The method of claim 1 , wherein the EMR comprises has a wavelength of about 172 nm.

14. The method of claim 1 , wherein the surface is substantially planar.

15. The method of claim 1 , wherein the surface comprises microstructures.

16. The method of claim 15 , wherein the microstructures comprise channels or wells.

17. The method of claim 1 , wherein the EMR is emitted from a lamp or a laser.

18. The method of claim 17 , wherein the lamp comprises an emission source in a shape of a cylinder or a flat panel.

19. The method of claim 1 , wherein the structure is a plate, tape, or belt.

20. A method for gene synthesis, the method comprising:
providing predetermined sequences for a plurality of oligonucleotides, wherein the plurality of oligonucleotides collectively encode for a plurality of genes;
providing a surface for oligonucleotide synthesis;
synthesizing the plurality of oligonucleotides from the surface, wherein each oligonucleotide extends from a different locus, and wherein the different loci are at least about 75% uniform when measured by calculating amplitude of signal variation for oligonucleotides extending from each locus divided by total signal intensity following white light illumination using an optical microscope; and
assembling the plurality of genes from the plurality of oligonucleotides.

21. The method of claim 20 , further comprising, prior to synthesizing:
providing the surface for oligonucleotide synthesis, wherein the surface comprises a first set of molecules, wherein each of the first set of molecules lacks a reactive group capable of binding to a nucleoside;
applying electromagnetic radiation (EMR) to predetermined regions of the surface, wherein the EMR comprises a wavelength from about 100 nm to about 300 nm, wherein application of the EMR results in removal of the first set of molecules at the predetermined regions, thereby defining loci for oligonucleotide extension.

Assignees

• Twist Bioscience Corp

Inventors

• Peck Bill James
• Indermuhle Pierre
• Marsh Eugene P
• Fernandez Andres
• Stern David

Classifications

• B01J2219/00432

  Photolithographic masks · CPC title

• B01J2219/00619

  using hydrophilic or hydrophobic regions · CPC title

• B01J2219/00711

  Light-directed synthesis · CPC title

• B01J2219/0061

  The surface being organic · CPC title

• B01J2219/00441

  using lasers · CPC title