Method and device for the integrated synthesis and analysis of analytes on a support

• US 20040043509A1
• Filed: 08/01/2003
• Published: 03/04/2004
• Est. Priority Date: 10/17/2000
• Status: Active Grant

• Current Assignee: Febit AG (FEBIT)
• Original Assignee: Febit AG (FEBIT)
• Inventors: Guimil, Ramon, Scheffler, Matthias, Stahler, Cord F., Heidbrede, Anke, Stahler, Peer F.
• Granted Patent: US 7,470,540 B2
• Time in Patent Office: Days
• US Class Current: 436/518
• 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 2219/00279 Features relating to reactor vessels
• CPC Class Codes: B01J 2219/00466 in a slurry
• CPC Class Codes: B01J 2219/00468 by manipulation of individual beads
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/0054 Means for coding or tagging the apparatus or the reagents
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00657 One-dimensional arrays
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• 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 60/14 for creating libraries
• CPC Class Codes: C40B 70/00 Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/163333 : Organic [e.g., chemical warfare agents, insecticides, etc.]
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A method for preparing receptor-coated particles is described which comprises the following steps:

a) providing a support body,

b) conducting a liquid containing particles into or onto the support body,

c) immobilizing the particles on at least one surface area of the support body,

d) conducting a liquid which contains receptors or receptor building blocks for synthesizing polymeric receptors over the immobilized particles,

e) coupling the receptors or receptor building blocks location- or/and time-specifically to the immobilized particles at in each case predetermined positions of the support body,

f) repeating, where appropriate, the steps (d) and (e), until the desired receptors have been synthesized on the immobilized particles at the in each case predetermined positions of the support body.

Claims

1. A method for integrated synthesis and analyte determination on a support, comprising the following steps:

a) providing a support body (2),

b) conducting a liquid containing particles (46) into or onto the support body (2),

c) immobilizing the particles (46) on at least one inner or/and outer surface (45) of the support body (2),

d) conducting a liquid which contains receptors or building blocks (A, B) for synthesizing polymeric receptors over the immobilized particles (46),

e) coupling the receptors or receptor building blocks (A, B) location- or/and time-specifically to the immobilized particles (46) at in each case predetermined positions of the support body (2),

f) repeating, where appropriate, the steps (d) and (e), until the desired receptors have been synthesized on the immobilized particles (46) at the in each case predetermined positions of the support body,

g) contacting the receptors with a sample containing the analyte(s) to be determined, and

h) determining the analyte(s) via binding to the receptors which are coupled to the immobilized particles (46).

2. The method as claimed in claim 1, characterized in that step (c) comprises permanent immobilizing.

3. The method as claimed in claim 2, characterized in that permanent immobilizing comprises a covalent chemical bond.

4. The method as claimed in claim 1, characterized in that step (c) comprises non-permanent immobilizing.

5. The method as claimed in claim 4, characterized in that non-permanent immobilizing comprises a magnetic, electrical or/and mechanical interaction.

6. The method as claimed in claim 4 or 5, furthermore comprising detaching the immobilized particles (46) from the support surface (45) and guiding the particles (46) out of the support body (2).

7. The method as claimed in any of the preceding claims, characterized in that synthesis and analyte determination are carried out in an integrated apparatus, the synthesis process or/and analyte determination process being monitored and controlled in any number of positions on the support.

8. The method as claimed in claim 7, characterized in that an integrated apparatus is used which comprises a programmable light source matrix (4), a detector matrix (6), a support body (2) which is preferably arranged between light source matrix and detector matrix and has at least some transparent regions, and means for supplying fluid into the support body and for discharging fluid from the support body.

9. The method as claimed in any of the preceding claims, characterized in that the analyte is removed from the support body again after the determination.

10. The method as claimed in any of the preceding claims, characterized in that a plurality of synthesis/analyte determination cycles are carried out, the receptors being synthesized for a subsequent cycle on the basis of the information from a preceding cycle.

11. The method as claimed in claim 10, characterized in that the receptors from the preceding cycle are extended for the subsequent cycle.

12. The method as claimed in claim 11, characterized in that a new support body having modified receptors compared with the preceding cycle is synthesized for the subsequent cycle.

13. The method as claimed in claim 12, characterized in that modification of the receptors comprises changing the sequence or/and eliminating negative receptors of the preceding cycle.

14. The method as claimed in any of the preceding claims, characterized in that a support body (2) having a multiplicity of channels (40) is used.

15. The method as claimed in any of the preceding claims, characterized in that a plurality of support bodies (2) are used for a single synthesis/analyte determination cycle.

19. An apparatus for carrying out the method as claimed in any of claims 1-15 or for carrying out at least one of the methods as claimed in claims 16, 17 and 18, comprising

a support body (2) which has at least one inner or/and one outer surface region (45) for attaching particles (46), in particular microbeads, whose surface has been functionalized,

means for supplying the particles (46) to the at least one surface region (45) of the support body (2),

a device (30) for immobilizing the particles (46) on the at least one surface region (45) of the support body (2),

means for supplying receptors or receptor building blocks A, B to particles (46) immobilized on the at least one surface region (45), and

means (4, 6, 8) for controlling location-and/or time-specific coupling of the receptors or receptor building blocks A, B to the immobilized particles (46) at in each case predetermined positions of the support body (2).

20. The apparatus as claimed in claim 19, in which the support body (2) has inner or/and outer channels, in particular microchannels (40), which have surface regions (45) for immobilizing the particles (46).

21. The apparatus as claimed in claim 19 or 20, in which the particles (46) are magnetic and in which the device (30) for immobilizing the particles (46) have means for generating a magnetic field B pervading the support body (2).

22. The apparatus as claimed in claim 21, in which the means (30) for generating a magnetic field comprise electromagnetic coils, in particular at least one Helmholtz coil pair, or/and at least one permanent magnet.

23. The apparatus as claimed in any of claims 19-22, in which the particles (46) are electrically charged or electrically polarized and in which the device (30) for immobilizing the particles (46) comprises means for generating an electric field E pervading the support body (2).

24. The apparatus as claimed in any of claims 19-23, in which the device (30) for immobilizing the particles (46) comprises pore openings in the at least one support-body surface region (45) for immobilizing the particles (46) and means for generating a suction action at the pore openings (47), which holds the particles in question (46) at the surface region (45′

).

25. The apparatus as claimed in claim 24, in which the surface regions provided with pore openings (47) are arranged in depressions (58) of microfluid channels (40a).

26. The apparatus as claimed in claim 24 or 25, in which the pore opening diameters are less than 90% of the smallest particle diameter, preferably less than 30% of the smallest particle diameter.

27. The apparatus as claimed in any of claims 19-26, in which the device (30) for immobilizing the particles (46) has been set up for the purpose of exerting in a controlled location-specific and space-resolved manner holding forces on particles in question in optionally different sections of the surface regions (46) of the support body (2).

28. The apparatus as claimed in any of claims 19-27, in which the means for controlling location-or/and time-specific coupling of the receptors or receptor building blocks (A, B) to the immobilized particles (46) has an illumination matrix (4) for generating illumination patterns which can be controlled in a programmable manner, and in which apparatus the support body (2) is, at least on one side, transparent for light from the illumination matrix (4) on the side of the illumination matrix (4), which faces the at least one surface region (45).

29. The apparatus as claimed in claim 28, which further has a light detection matrix (6) for optical monitoring of particle immobilization or/and of synthesis processes or/and of analyte determination processes in the support body (2).

30. The apparatus as claimed in any of claims 19-29, in which the particles (46) have diameters of essentially the same size and in which the particle diameter is less than 50 μ

m, in particular less than 10 μ

m.

16. A method for preparing receptor-coated particles for determining analytes, comprising the following steps:

a) providing a support body (2),

b) conducting a liquid containing particles (46) into or onto the support body (2),

c) immobilizing the particles (46) on at least one inner or/and outer surface (45) of the support body (2),

d) conducting a liquid which contains receptors or building blocks (A, B) for synthesizing polymeric receptors over the immobilized particles (46),

e) coupling the receptors or receptor building blocks (A, B) location- or/and time-specifically to the immobilized particles (46) at in each case predetermined positions of the support body (2),

f) repeating, where appropriate, the steps (d) and (e), until the desired receptors have been synthesized on the immobilized particles (46) at the in each case predetermined positions of the support body.

17. A method for determining one or more analytes on a support, comprising the following steps:

a) providing a support body (2) having at least one inner or/and outer surface (45), where particles (46) with receptors coupled thereto are immobilized at in each case predetermined positions of the surface, said particles (46) carrying different receptors in different regions,

g) contacting the support body (2) with a sample containing the analyte(s) to be determined, and

h) determining the analyte(s) via binding to the receptors coupled to the immobilized particles (46).

18. A method for determining one or more analytes on a support, comprising the following steps:

a) providing a support body (2) having at least one inner or/and outer surface (45), where particles (46) with receptors coupled thereto are immobilized to the surface (45), with a plurality of coded particle species having in each case different receptors being used and the immobilization comprising a magnetic or/and electrical interaction,

g) contacting the support body or the receptors with a sample containing the analyte(s) to be determined, and p1 h) determining the analyte(s) via binding to the receptors coupled to the immobilized particles (46).