Binding assays using magnetically immobilized arrays

• US 20030040129A1
• Filed: 07/26/2002
• Published: 02/27/2003
• Est. Priority Date: 08/20/2001
• Status: Abandoned Application

• Current Assignee: Haresh P. Shah
• Original Assignee: Haresh P. Shah
• Inventors: Shah, Haresh P.
• Application Number:
  US10/206,721
• Publication Number:
  US 20030040129A1
• Time in Patent Office: Days
• US Class Current: 436/526
• 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: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/553 Metal or metal coated
• CPC Class Codes: G01N 35/0098 involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C

Abstract

Systems and methods for preparing and using magnetic particle arrays are provided. In conventional assays, a target molecule is immobilized in a particular array position on the surface of the substrate by chemically conjugating the molecule to the surface of the substrate. According to embodiments of the present invention, target biomolecules are immobilized magnetically rather than chemically. Accordingly, the target molecules are chemically conjugated to the surface of a magnetic particle, and it is the magnetic particles that are positioned in an array by printing (or spotting) the magnetic particles onto the surface of a magnetic array substrate. The array is exposed to a solution of probe molecules (analytes) having detector labels, and the positions in the array where complementary target binds to probe are recorded. Such magnetic particle arrays may be used in a variety of applications, including drug screening, nucleic acid sequencing, mutation analysis, medical diagnosis, and immunoassay analysis. The magnetic particle array may be The advantages of a magnetic particle array, which may be configured with a holder having microfluidic channels to deliver sample to the array to comprise a magnetic array biochip, include reduced assay variability, enhanced flexibility, lower cost and higher throughput.

Claims

1. A magnetic particle array for performing biological and chemical assays, the magnetic particle array comprising:

at least one magnetic layer comprising a predetermined spatial distribution of spaced-apart magnetized spots;

a plurality of magnetic particles immobilized to said magnetic spots;

wherein at least a plurality of said particles comprise different molecules.

2. The magnetic particle array of claim 1 further comprising:

an array support layer to which said magnetic layer is attached.

3. The magnetic particle array of claim 2, further comprising a biochip holder for supporting

the magnetic particle array, wherein the biochip holder contains at least one fluidic channel for transporting a reagent(s) to the magnetic particle array.

4. The magnetic particle array of claim 3, wherein the biochip holder further comprises an inlet port for introducing a reagent to the magnetic particle array via the fluidic channel.

5. The magnetic particle array of claim 3, wherein the biochip holder further comprises a reaction chamber adjacent to the magnetic particle array.

6. The magnetic particle array of claim 1, wherein said different molecules comprise one or more molecules from group consisting of, but not limited to, chemical molecules, polymers, biopolymers, proteins, peptides, cells, enzymes, substrates, antibody, antigen, bacteria, virus, hapten, drugs, receptors, recombinant molecules, DNA, RNA, PNA, Poly A, oligonucleotides, carbohydrates, hormones, metal sol, metal chelates, dyes, particulates, label moiety such as fluorescent or luminescent molecules, member of specific binding reaction and the like.

7. The magnetic particle array of claim 2, wherein the array support layer is made from a polymer or combination of polymers, plastic,, silicon, resin, polysaccharide, silica, hydrogel, carbon, metal, inorganic glasses, membrane, porous or non-porus material, and paper-based material.

8. The magnetic particle array of claim 2, wherein the surface of the array support layer contains a flexible or rigid structure(s) selected from the group consisting of:

planar, irregular, a well, grooves, multi-well plate, dish, screen, mesh, depression, elevation, trench, chamber, nanowells, array of pits, strips, dipstick, fluidic network, capillary, porous film, non-porous film, gel and channels.

9. The magnetic particle array of claim 2, wherein the array support layer has a surface treated or coated, derivatized or modified with chemical or polymer or biopolymer.

10. The magnetic particle array of claim 1, wherein the at least one magnetic layer comprises a material selected from the group consisting of:

, but not limited to, a fine magnetic powder loaded into a thermoplastic binder;

a bonded material comprising neodymium, iron, and boron;

;

a sheet of plastic material impregnated with a ferromagnetic material; and

a sheet of synthetic resin material having mixed therein magnetic powder particles, neodymium-iron-boron, samarium-cobalt, barium ferrite, and strontium ferrite.

11. The magnetic particle array of claim 1, wherein the at least one magnetic layer has a surface treated or coated, derivatized or modified with chemical or polymer or biopolymer.

12. The magnetic particle array of claim 1, wherein said magnet layer has one dimension in the range of from about 0.0001 to 0.5 inches.

13. The magnetic particle array of claim 1, wherein said magnetic layer has a magnetic field strength in the range of about 100 to 15,000 Gauss.

14. The magnetic particle array of claim 1, wherein said magnetic layer is patterned permanent magnet.

15. The magnetic particle array of claim 1, wherein said magnetic layer is electromagnetic.

16. The magnetic particle array of claim 1, wherein said magnetic layer, is produced by any number of methods including;

extrusion, calendering, injection molding, compression molding, printing, spin coating, chemical or vapor deposition, sputtering and combinations thereof.

17. The magnetic particle array of claim 1, wherein said magnetic particles comprise a magnetic core surrounded by an organic or polymer coating.

18. The magnetic particle array of claim 17, wherein the magnetic core of the magnetic particles is selected from the group consisting of one or more metals, metal oxides, metal salts, metal hydroxides, alloys of metals, organometallic compounds, and mixtures thereof.

19. The magnetic particle array of claim 1, wherein magnetic particles are swellable

20. The magnetic particle array of claim 1, wherein magnetic particles are non-swellable

21. The magnetic particle array of claim 1, wherein the mean diameter of the magnetic particles ranges from about 0.05 to 1,000 μ

m.

22. The magnetic particle array of claim 1, wherein the areal density of magnetic particle array spots on the surface of the array support layer ranges from about 1 to 100 spots per mm².

23. The magnetic particle array of claim 1, wherein the number of magnetic particles per array spot ranges from about 1 to 10⁶.

24. The magnetic particle array of claim 1, wherein the total number of spots in the array ranges up to about 100,000.

25. The magnetic particle array of claim 1, wherein the largest dimension of each of the array spot ranges from about 1 um to 5 mm.

26. The magnetic particle array of claim 1, wherein the pattern of spots in the array is selected from the group consisting of orthogonally organized rows and columns, grids, curvilinear rows across the substrate surface, concentric circles, concentric semi-circles, and simple rows of lines.

27. The magnetic particle array of claim 1, wherein said different molecules are attached to magnetic particles prior to forming magnetic particle array.

28. The magnetic particle array of claim 1, wherein said different molecules are attached to magnetic particles after forming magnetic particle array.

29. The magnetic particle array of claim 1, wherein each magnetic array spot contains magnetic particles coated with identical or mixture of different molecules.

30. The magnetic particle array of claim 1, wherein each array spot contains one or more type of magnetic particles, wherein each particle type is distinguishable

31. A magnetic particle array as set forth in claim 1, comprises at least about 1 to 96 or more substantially identical, spatially discrete regions, each region comprising, magnetic particle array from about 2 to 500 spots.

32. A method of making the said magnetic particle array comprising;

to a magnetic array layer comprised of plurality of magnetized spots,

dispensing or microspotting reagent containing magnetic particles on the surface of the magnetic layer in spatially defined and physically addressable manner until the said magnetic array is formed.

33. A magnetic particle array comprising;

a magnet layer having individual permanently magnetized spots at a density in the range of about 1 to 100 spots per mm², each of said spots having a maximum planar dimension in the range of about 1 um to 5 mm;

immobilized at a plurality of said spots magnetic particles having diameter in the range of about 0.005 to 1,000 um, a plurality of said magnetic particles having different molecules of interest conjugated to said magnetic particles.

34. A method of using a magnetic particle array to perform an assay employing magnetic particles to which target molecules have been conjugated and a magnetic array substrate having spaced-apart magnetized spots, the method comprising:

transferring the said magnetic particles onto the surface of the magnetic array substrate with said magnetic particles being immobilized at said spaced apart magnetized spots to form a magnetic particle array with the said magnetic particles in a predetermined organization; and

adding a solution comprising a composition of interest to said magnetic particle array; and

determining any interaction between said composition of interest and said target molecule.

35. The said method of claim 34, wherein the step of transferring the magnetic particles onto the surface of the magnetic array substrate further comprises loading the magnetic particles into the tip of the pipette, and dispensing the magnetic particles at a predetermined location as a spot in the array.

36. The method of claim 34, wherein the composition of interest further comprises a label or a labeled complementary binding molecule is added to bind to said composition of interest.

37. The said method of claim 34, wherein the step of transferring the magnetic particles onto the surface of the magnetic array substrate further comprises transferring the magnetic particles with an electromagnetic transfer system

38. A system for screening reaction between compositions of interest, said system comprising;

vessels organized in a spatial arrangement containing magnetic particles, wherein a plurality of said vessels contain magnetic particles having different composition of interest;

a magnetic layer comprising magnetized spots spatially organized in conformance with said vessel spatial arrangement; and

a magnetic transfer device for simultaneously retrieving magnetic particles from each of the said vessels and transferring said magnetic particles to said magnetic layer in the spatial organization of said vessels.

39. A system according to claim 38, wherein said vessels are microtiter well plates

40. A system according to claim 38, further comprising a data processing unit

41. A system according to claim 38, wherein said magnetic particles have a diameter in the range of about 0.05 to 1000 um and said spots are at a density in the range of about 1 to 100 spots per mm², each of said spots having a maximum planar dimension in the range of about 1 um to 5 mm.