Solvent for biopolymer synthesis, solvent microdroplets and methods of use

• US 6,384,210 B1
• Filed: 03/13/2000
• Issued: 05/07/2002
• Est. Priority Date: 03/20/1997
• Status: Expired due to Term

• Current Assignee: University of Washington
• Original Assignee: University of Washington
• Inventors: Blanchard, Alan P.
• Primary Examiner(s): Whisenant, Ethan C.
• Application Number:
  US09/381,487
• Time in Patent Office: 785 Days
• Field of Search: 536/25.3, 536/23.1, 536/24.3, 422/129, 422/131, 347/1, 347/85, 347/7, 347/40, 347/107
• US Class Current: 536/25.3
• 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 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00475 Sheets
• CPC Class Codes: B01J 2219/00488 by rotation of the reaction vessels
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00536 in the shape of disks
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00675 In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01J 2219/00695 Synthesis control routines, e.g. using computer programs
• CPC Class Codes: B01J 2219/00704 : integrated with the reactor apparatus
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B41J 2/01 : Ink jet
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07H 19/00 : Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
• 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
• CPC Class Codes: C07K 1/02 : in solution C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/04 : on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/14 : Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support

Abstract

The present invention provides a method of biopolymer, esp. oligonucleotide, synthesis. The method consists of coupling a first nucleotide to a second nucleotide in a high surface tension solvent. The invention also provides microdroplets of a soln. comprising a solvent having a b.p. of 150 degree. C. or above, a surface tension of 30 dynes/cm or above, and a viscosity of 0.015 g/(cm)(sec), e.g., propylene carbonate. Such microdroplets are useful for the synthesis of chem. species, particularly biopolymers such as oligonucleotides and peptides, as well as arrays of chem. species. An automated system for oligonucleotide synthesis is described, which comprises delivery of microdroplets by inkjet technol. and computer control of the process. The high surface tension solvent used is selected for compatibility with the inkjet technol.

Claims

1. An inkjet print head comprising a piezoelectric pump, wherein said print head contains a solvent having a boiling point of 150°

C. or above, a surface tension of 30 dynes/cm or above, and a viscosity of 0.015 g/(cm)(sec) or above, which inkjet printhead does not contain a solvent having a hydroxyl group.

2. The inkjet print head of claim 1, wherein the solvent has the formula (I):

wherein

A=O or S;

X=O, S or N(C₁-C₄ alkyl);

Y=O, S, N(C₁-C₄alkyl) or CH₂; and

R=C₂-C₂₀ straight or branched chain alkyl.

3. The inkjet print head of claim 2, wherein said solvent is selected from the group consisting of:

N-methyl-2-pyrrolidine;

2-pyrrolidine;

propylene carbonate;

γ

-valerolactone;

6-caprolactam;

ethylene carbonate;

γ

-butyrolactone;

δ

-valerolactone;

1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

ethylene trithiocarbonate; and

1,3-dimethyl-2-imidazolidinone.

4. The inkjet print head of claim 3, wherein the solvent is propylene carbonate.

5. The inkjet print head of claim 1, which does not contain a solvent having a primary or secondary amino group.

6. The inkjet print head of claim 1, which does not contain a solvent having a sulfhydryl group.

7. The inkjet print head of claim 1, which does not contain a solvent having a carboxyl group.

8. The inkjet print head of claim 1, which does not contain a solvent having an anhydride group.

9. An inkjet print head containing a solution comprising a solvent having a boiling point of 150°

C. or above, a surface tension of 30 dynes/cm or above, and a viscosity of 0.015 g/(cm)(sec) or above, and a nucleoside or an activated nucleoside.

10. The inkjet print head of claim 9, wherein the solvent has the formula (I):

wherein

A=O or S;

X=O, S or N(C₁-C₄ alkyl);

Y=O, S, N(C₁-C₄ alkyl) or CH₂; and

R=C₂-C₂₀ straight or branched chain alkyl.

11. The inkjet print head of claim 10, wherein the solvent is selected from the group consisting of:

N-methyl-2-pyrrolidone;

2-pyrrolidone;

propylene carbonate;

γ

-valerolactone;

6-caprolactam;

ethylene carbonate;

γ

-butyrolactone;

δ

-valerolactone;

1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

ethylene trithiocarbonate; and

1,3-dimethyl-2-imidazolidinone.

12. The inkjet print head of claim 11, wherein the solvent is propylene carbonate.

13. An inkjet print head containing a solution comprising propylene carbonate and a nucleoside having a phosphodiester, phosphotriester, phosphate triester, H-phosphonate or phosphoramidite group.

14. An inkjet print head containing a solution comprising a solvent having a boiling point of 150°

C. or above, a surface tension of 30 dynes/cm or above, and a viscosity of 0.015 g/(cm)(sec) or above, and an amino acid.

15. The inkjet print head of claim 14, wherein the solvent has the formula (I):

wherein

A=O or S;

X=O, S or N(C₁-C₄ alkyl);

Y=O, S, N(C₁-C₄ alkyl) or CH₂; and

R=C₂-C₂₀ straight or branched chain alkyl.

16. The inkjet print head of claim 15, wherein the solvent is selected from the group consisting of:

N-methyl-2-pyrrolidone;

2-pyrrolidone;

propylene carbonate;

γ

-valerolactone;

6-caprolactam;

ethylene carbonate;

γ

-butyrolactone;

δ

-valerolactone;

1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

ethylene trithiocarbonate; and

1,3-dimethyl-2-imidazolidinone.

17. The inkjet print head of claim 16, wherein the solvent is propylene carbonate.

18. An inkjet print head containing a solution comprising a solvent having a boiling point of 150°

C. or above, a surface tension of 30 dynes/cm or above, and a viscosity of 0.015 g/(cm)(sec) or above, and a catalyst, which inkjet printhead does not contain a solvent having a hydroxyl group.

19. The inkjet print head of claim 18, wherein the solvent has the formula (I):

wherein

A=O or S;

X=O, S or N(C₁-C₄ alkyl);

Y=O, S, N(C₁-C₄ alkyl) or CH₂; and

R=C₂-C₂₀ straight or branched chain alkyl.

20. The inkjet print head of claim 19, wherein the solvent is selected from the group consisting of:

N-methyl-2-pyrrolidone;

2-pyrrolidone;

propylene carbonate;

γ

-valerolactone;

6-caprolactam;

ethylene carbonate;

γ

-butyrolactone;

δ

-valerolactone;

1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

ethylene trithiocarbonate; and

1,3-dimethyl-2-imidazolidinone.

21. The inkjet print head of claim 20, wherein the solvent is propylene carbonate.

22. The inkjet print head of any of claims 18-21, wherein said catalyst is selected from the group consisting of 5-methylthiotetrazole, tetrazole, 5-ethylthiotetrazole and dicyclohexylcarbodiimide.

23. An inkjet print head containing a solution comprising a solvent having a boiling point of 150°

C. or above, a surface tension of 30 dynes/cm or above, and a viscosity of 0.015 g/(cm)(sec) or above, and a catalyst, wherein said catalyst is selected from the group consisting of 5-methylthiotetrazole, tetrazole, 5-ethylthiotetrazole and dicyclohexylcarbodiimide.

24. The inkjet print head of claim 23, wherein the solvent has the formula (I):

wherein

A=O or S;

X=O, S or N(C₁-C₄ alkyl);

Y=O, S, N(C₁-C₄ alkyl) or CH₂; and

R=C₂-C₂₀ straight or branched chain alkyl.

25. The inkjet print head of claim 24, wherein the solvent is selected from the group consisting of:

N-methyl-2-pyrrolidone;

2-pyrrolidone;

propylene carbonate;

γ

-valerolactone;

6-caprolactam;

ethylene carbonate;

γ

-butyrolactone;

δ

-valerolactone;

1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

ethylene trithiocarbonate; and

1,3-dimethyl-2-imidazolidinone.

26. The inkjet print head of claim 25, wherein the solvent is propylene carbonate.

27. An inkjet print head containing a solvent selected from the group consisting of:

2-pyrrolidone;

propylene carbonate;

γ

-valerolactone;

6-caprolactam;

ethylene carbonate;

γ

-butyrolactone;

δ

-valerolactone;

1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

ethylene trithiocarbonate; and

1,3-dimethyl-2-imidazolidinone,

which inkjet printhead does not contain a solvent having a hydroxyl group.

28. An inkjet print head containing propylene carbonate, which inkjet printhead does not contain a solvent having a hydroxyl group.