Patterned polymer synthesis

• US 20020168669A1
• Filed: 03/26/2002
• Published: 11/14/2002
• Est. Priority Date: 03/26/2001
• Status: Abandoned Application

• Current Assignee: Linden Technologies Incorporated
• Original Assignee: Linden Technologies Incorporated
• Inventors: Huang, Tai-Nang
• Application Number:
  US10/107,556
• Publication Number:
  US 20020168669A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00385 Printing
• CPC Class Codes: B01J 2219/00459 Beads
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00614 Delimitation of the attachment areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids

Abstract

An array of chemical compounds can be produced by the electrostatic deposition of its components onto a substrate. The subunit building blocks are coupled to the chemical groups on the substrate to synthesize a complex compound. By localizing the electrostatic deposition, different building blocks can be coupled at different positions on the substrate. Thus, a diverse and addressable set of chemical compounds is produced on the substrate to form an array of chemical compounds, e.g., of biological polymers. One application of this concept is the production of an oligonucleotide array. Other concepts provided here can be used in combination with the electrostatic deposition method or with other chemical synthetic methods. The invention provides, in part, methods of dispensing the nucleic acid subunits as a dry composition (e.g., a particulate composition) for the in-situ synthesis of nucleic acid polymers.

Claims

1. A method of constructing a nucleic acid array, the method comprising:

determining a plurality of layer maps from a set of nucleic acid sequences, each layer map corresponding to a particular register and indicating discrete addresses at which corresponding nucleic acid subunits are to be coupled;

for each register, directing a plurality of applicator units that each are supplied with a different nucleic acid subunit to each transfer the respective nucleic acid subunit to a substrate at the discrete addresses indicated by the layer map for the respective nucleic acid subunit;

coupling the transferred nucleic acid subunits to a reactive terminus to form an array of nucleic acids.

2. The method of claim 1 wherein the reactive terminus is on the surface of the substrate.

3. The method of claim 1 wherein the reactive terminus is on the surface of a second substrate, and the nucleic acid subunits are transferred from the first substrate to the second substrate.

4. The method of claim 1 wherein the nucleic acid subunits are supplied as particles of less than 50 μ

m diameter.

5. The method of claim 1 wherein each applicator unit comprises a photoreceptor, a light unit that selectively alters the charge at discrete positions on the photoreceptor, and an agitator that agitates a toner composition comprised of the nucleic acid subunit particles and carrier beads.

6. The method of claim 1 wherein the directing comprises sending the layer map for the register to each of the applicator units.

7. The method of claim 1 wherein the directing comprises generating a plurality of image maps for the register, the image maps indicating discrete addresses at which a given nucleic acid subunit is to be coupled, and sending each of the image maps of the plurality to the applicator unit that is supplied with the corresponding nucleic acid subunit.

8. The method of claim 7 wherein the image maps are pixilated.

9. The method of claim 1 wherein the determining is effected by a processor.

10. The method of claim 9 wherein the processor is in signal communication with each of the applicator units of the plurality.

11. The method of claim 1 wherein the applicator units comprise a common belt that rotates between the applicator units, and the belt includes a photoreceptive surface.

12. The method of claim 3 wherein the first substrate is flexible.

14. The method of claim 13, wherein the surface is the surface of an interim substrate and the method comprises transferring the particles from the surface of the interim substrate to the solid support.

15. The method of claim 14, wherein a different interim substrate is used for each repetition of 1) and 2).

16. The method of claim 13 wherein the coupling comprises contacting the particles to an activator compound.

17. The method of claim 16 wherein the activator compound is selected from the group consisting of:

tetrazole, 5-(p-nitrophenyl)-1H-tetrazole, 5-ethylthio-1H-tetrazole, 4,5-dichloroimidazole, benzimidazolium triflate, or 4,5 dicyano-imidazole.

18. The method of claim 13 wherein a cycle of a) to d) comprises conveying the substrate between each of four applicator units, each applicator unit comprising a photoreceptor and a source of the respective nucleotide particles.

19. The method of claim 13 wherein the substrate is flexible.

20. The method of claim 19 wherein the substrate comprises paper, Mylar, cellulose, polyvinylchloride, and/or polycarbonate.

13. A method of synthesizing nucleic acids, the method comprising:

1) repeating for each of four nucleotide bases, adenine, guanine, cytosine, and thymidine,

a) triboelectrically charging particles of the respective nucleotide base, the base including a protecting group;

b) selectively irradiating a photoreceptor to generate a patterned region with defined electrostatic charge;

c) contacting the charged nucleotide particles to the photoreceptor to attach the particles to the photoreceptor in the patterned region; and

d) transferring the nucleotide particles from the photoreceptor to a surface;

2) coupling the transferred particles to terminal groups on a solid support; and

3) repeating

     1) and

     2) to produce a plurality of different nucleic acid sequences on the solid support.

21. A method comprising:

combining toner particles less than 50 microns diameter and charged carrier beads to generate a developer particle complexes, the toner particles including a compound having a reactive group and a protecting group;

loading the toner particles from the developer particles complexes in a loading zone onto a surface region of a photoreceptor, the surface region being selectively charged by illumination;

positioning the photoreceptor to displace the surface region from the loading zone into proximity to or contact with a substrate;

transferring the protected chemical compound to the substrate.

22. The method of claim 21 wherein the compound is a monomeric subunit of a biopolymer.

23. The method of claim 22 wherein the compound is a nucleotide that includes a protecting group.

24. The method of claim 23 wherein the nucleotide compound also includes a phosphorous activating group.

25. The method of claim 24 further comprising contacting a solvent containing an activator compound to the substrate, wherein the activator compound couples the reactive group of the protected chemical compound to an immobilized group on the substrate.

26. The method of claim 23 wherein the activator compound is 5-ethylthio-1H-tetrazole or tetrazole.

27. The method of claim 21 wherein the transferring comprises generating an electric field.