Method of spotting probes on a solid support

• US 7,994,297 B2
• Filed: 04/20/2006
• Issued: 08/09/2011
• Est. Priority Date: 08/01/1997
• Status: Expired due to Fees

• Current Assignee: Canon Kabushiki Kaisha (Canon Inc.)
• Original Assignee: Canon Kabushiki Kaisha (Canon Inc.)
• Inventors: Suzuki, Tomohiro, Okamoto, Tadashi, Yamamoto, Nobuko
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US11/407,201
• Publication Number:
  US 20060177868A1
• Time in Patent Office: 1,937 Days
• Field of Search: None
• US Class Current: 536/23.1
• 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 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel 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 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• 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 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00729 : Peptide nucleic acids [PNA]
• CPC Class Codes: B01L 2200/143 : Quality control, feedback systems
• CPC Class Codes: B01L 2400/0442 : thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 3/0268 : using pulse dispensing or spraying, eg. inkjet type, piezo actuated ejection of droplets from capillaries
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 : Compounds covalently bound to a solid support
• 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: C12N 11/14 : Enzymes or microbial cells immobilised on or in an inorganic carrier
• CPC Class Codes: C12N 15/11 : DNA or RNA fragments; Modified forms thereof DNA or RNA not used in recombinant technology, C07H21/00; Non-coding nucleic acids having a biological activity
• CPC Class Codes: C12N 2310/351 : Conjugate
• CPC Class Codes: C12Q 1/00 : Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C40B 30/04 : by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, 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
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/68 : involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures

Abstract

Provided is a method of spotting a probe densely and efficiently on a surface of a solid support. A liquid containing a probe is attached to a solid support as droplets to form spots containing the probe on the solid support by an ink jet method.

Claims

1. A method of producing a probe array, said method comprising the steps of:

spotting in a spot on a surface of a solid support by an ink-jet method a plurality of single-stranded nucleic acid probes, which are in an aqueous solution and

effecting covalent bonding of the probes to the surface of the solid support through the intermediary of a substance, which substance is derived from N-(6-maleimidocaproyloxy) succinimide, the covalent bonding being effected via a reaction between a thiol group of a probe, the probe being one of the probes, with a maleimido group of the substance, with the reaction forming the covalent bonding between the thiol group of the probe and the maleimido group of the substance occurring after the spotting step,

wherein in the ink-jet method, thermal energy is applied to a liquid to eject the liquid from a nozzle as a flying liquid droplet.

2. A method according to claim 1, wherein the solid support has a homogeneous surface property.

3. A method according to claim 1, wherein the probes are single-stranded DNA probes.

4. A method according to claim 1, wherein the probes are single-stranded RNA probes.

5. A method according to claim 1, wherein the probes are single-stranded PNA probes.

6. A method according to claim 5, wherein the probes are single-stranded PNA probes and contain a cysteine residue on an N-terminus side.

7. A method according to claim 1, wherein the distance between adjacent spots is not smaller than a maximum width of the spots.

8. A method according to claim 1, wherein the density of the spots is 10,000 per square inch or higher.

9. A method according to claim 1, wherein the surface is a flat surface.

10. A method according to claim 1, wherein the single-stranded nucleic acid probes are the same.

11. A method according to claim 1, wherein at least two spots are arranged on the surface, wherein a plurality of first single-stranded nucleic acid probes are arranged in one spot of the at least two spots, wherein a plurality of second single-stranded nucleic acid probes are arranged in another spot of the at least two spots, and wherein the first probes are different from the second probes.

12. A method according to claim 1, comprising the further steps of:

providing a plurality of liquids containing a plurality of probes, which are different from each other but are equal in mol concentration, the probes having a thiol group at the terminal end thereof for binding to a malemide group on the solid support; and

supplying an equal quantity per each of the liquids onto the surface of the solid support by using an ink-jet method in which droplets of the liquids are ejected from a nozzle by applying heat energy to the liquids.

13. A method according to claim 1, wherein the spot is about 4-50 picoliters.

14. A method according to claim 13, wherein the spot has a diameter of about 70-100 μ

m.

15. A method according to claim 13, wherein the probes have a concentration in the aqueous solution of 0.05-500 μ

M, and the solution has a viscosity of 1-15 cps and a surface tension of 30 dyn/cm or higher.

16. A method according to claim 13, wherein the probes have a concentration in the aqueous solution of 2-50 μ

M, and the solution has a viscosity of 1-5 cps and a surface tension of 30-50 dyn/cm.

17. A method according to claim 13, wherein in the ink-jet method, one liquid ejection is followed by plural idle ejections.

18. A method according to claim 13, wherein in the ink-jet method, a plurality of ink jet heads are used.

19. A method according to claim 1, wherein the spot has a diameter of about 20-100 μ

m.

20. A method of producing a probe array having a solid support with a plurality of probe spots on the surface of the solid support, which comprises the steps of:

i.) providing a plurality of liquids containing a plurality of probes, which are different from each other but are equal in mol concentration, the probes having a first functional group at the terminal end thereof for binding to the solid support;

ii.) providing a solid support of which the surface has a second functional group for binding to the first functional group; and

iii.) supplying an equal quantity per each of the liquids onto the surface of the solid support by using an ink-jet method in which droplets of the liquids are ejected from a nozzle by applying heat energy to the liquids, whereby a plurality of probe spots are formed and a covalent bonding is effected between the first functional group and the second functional group,

wherein the probe spots have different nucleic acids, and each of the nucleic acid probes is for specifically binding to a target substance.

21. A method according to claim 20, wherein the first functional group is a thiol group and the second functional group is a maleimide group.