Device and method for real-time detection of molecular accumulations and/or monitoring the production process of a molecular microarray

• US 9,675,954 B2
• Filed: 05/23/2013
• Issued: 06/13/2017
• Est. Priority Date: 05/23/2012
• Status: Active Grant

• Current Assignee: Biocopy GmbH
• Original Assignee: Albert Ludwigs University of Freiburg
• Inventors: Roth, Guenter
• Primary Examiner(s): Flinders, Jeremy C
• Application Number:
  US14/402,693
• Publication Number:
  US 20150209752A1
• Time in Patent Office: 1,482 Days
• 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 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00576 fluorophore
• 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 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00693 Means for quality control
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides

Abstract

The invention relates to a method for producing a microarray, wherein the production of this array is detected in real time from the accumulation of the product molecules being produced. The invention further relates to a microarray produced by this method, and to a device for the real-time detection of molecular accumulations on an array surface during the production of microarrays.

Claims

1. A method for producing a microarray, comprising

a) providing a first original array comprising a first support surface comprising reference spots that comprise reference molecules, to which at least one template molecule is bound,

b) providing a second support surface (receiver surface), which is brought into a spatial orientation relative to the first support surface,

c) providing a liquid, containing an enzymatic and/or chemical reaction system, between the first and second support surfaces, so that regions of the two surfaces are in contact with one another via the liquid,

d) producing product molecules using the template molecule and the enzymatic and/or chemical reaction system from said reference molecules,

e) transferring the product molecules to the second support surface via the liquid between the first and second support surfaces, wherein a correlation exists between a binding site of the template molecule to the first support surface and an accumulation of the corresponding product molecules on the second support surface, and

f) detecting the accumulation of the product molecules on the receiver surface in real time,

wherein a production of the product molecules according to steps e) and f) is detected in real time, and the producing of d), the transferring of e) and the detecting of f) are terminated as soon as a predefined reference spot size has been reached.

2. The method according to claim 1, wherein

a gap exists between the first and second support surface, which is filled with the liquid before, after or during the spatial orientation of the first and second support surfaces.

3. The method according to claim 1, wherein

the detecting in real time involves a label-free method, that detects the mass growth, the change in the refractive index, or the change in optical, magnetic, electric and/or electromagnetic properties.

4. The method according to claim 1, wherein

the detecting in real-time involves a method that uses a label.

5. The method according to claim 1, wherein

in the detecting in real-time the receiver surface serves as a waveguide.

6. The method according to claim 1, wherein said detecting comprises

a method selected from a group consisting of physical methods, electrical methods, chemical methods, fluorescence measurements, luminescence measurements, spectroscopy, absorption measurements, polarization measurements, reflectometric interference spectroscopy, imaging reflectometric interference spectroscopy, surface plasmon resonance spectroscopy, biacore, ellipsometry, polarization anisotropy, turbidity measurement, pH measurement, measurement of available H+ions, measurement with excited surface fluorescence, internal total reflection fluorescence microscopy, measurement using quenched surface fluorescence, measurements using evanescent field excitation, measurement using Raman spectroscopy and combinations thereof.

7. The method according to claim 1, wherein

the microarray that is produced is a DNA array, an RNA array, a protein array and/or an array of synthetic molecules.

8. The method according to claim 1, wherein

the template molecule is selected from a group consisting of DNA, RNA, genomic DNA, cloned DNA fragments, plasmid DNA, cDNA, PCR products, synthesized DNA products, synthesized DNA oligonucleotides, m RNA, synthesized RNA and combinations thereof.

9. The method according to claim 1, wherein

the original array is a DNA array and or a RNA array.

10. The method according to claim 1, wherein

the enzymatic reaction system according to c) comprises DNA polymerase, RNA polymerase and/or reverse transcriptase.

11. The method according to claim 1, wherein

the chemical reaction system comprises a cell-free mixture, which produces from DNA and/or RNA corresponding proteins.

12. The method according to claim 1, wherein

the detecting is used for monitoring a production process of the microarray.

13. The method according to claim 1, wherein

the original array is a planar array that contains DNA or RNA.

14. The method according to claim 1, wherein

following production of the microarray, a binding or interaction is also measured in f).

15. The method according to claim 13, wherein the planar array is a sequencing chip.