Static micro-array of biological or chemical probes immobilised on a support by magnetic attraction
AssigneeCommissariat a l'Energie Atomique
InventorsNicolas Ugolin, Gerard Marguerie De Rotrou, Thierry Kortulewski, Olivier Alibert, Diana Le Roux
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Abstract
The invention relates to an organized array of biological or chemical probes bonded to a support by magnetic coupling using a fixing vector. The support may include a permanent magnet or present electrically-induced magnetization. The fixing vectors may be constituted by magnetic beads functionalized to be capable of bonding specifically to one particular type of biological probe.
Claims30
1/ An organized array of biological or chemical probes bound to a support by magnetic coupling using a fixing vector.
2/ An array according to claim 1, fixed to a support that is optically neutral.
3/ An array according to claim 1 or claim 2, fixed to a support that includes a permanent magnet.
4/ An array according to claim 1 or claim 2, fixed to a support that presents electrically-induced magnetism.
5/ An array according to any one of claims 1 to 4, fixed to a support presenting structured magnetism.
6/ An array according to any one of claims 1 to 5, in which the magnetic field is perpendicular or parallel to the plane of the support.
7/ An array according to any one of claims 1 to 6, in which the fixing vector is paramagnetic, superparamagnetic, or ferromagnetic.
8/ An array according to claim 7, in which the fixing vector is a bead of latex or of polysaccharides including particles of iron oxide.
9/ An array according to claim 7 or claim 8, in which the fixing vector is a multipolar bead.
10/ An array according to claim 7 or claim 8, in which the fixing vector is capable of orienting itself in a magnetic field.
11/ An array according to claim 10, in which the fixing vector is itself magnetized.
12/ An array according to any one of claims 7 to 11, in which the fixing vector has a diameter lying in the range 1 nm to 500 μm, and preferably in the range 0.5 μm to 5 μm.
13/ An array according to any one of claims 1 to 12, in which the fixing vector carries elements capable of bonding specifically to a biological target.
14/ An array according to claim 13, in which the elements capable of bonding specifically to a biological target carried by the fixing vector are selected from the group comprising:
- immunoglobulins, antigens, membrane receptors, membrane receptor ligands, avidin, streptavidin, poly-T or poly-U oligonucleotides.
15/ An array according to any one of claims 1 to 14, in which each probe is deposited as a spot of diameter lying in the range 10 μm to 1 mm, and preferably in the range 50 μm to 200 μm.
16/ An array according to any one of claims 1 to 15, in which each spot has 105 to 1010 probes, and preferably 108 to 1010 probes.
17/ An array according to any one of claims 1 to 16, having a density of 1 to 100,000 spots per cm2, and preferably of 10 to 1000 spots per cm2.
18/ A method of manufacturing an organized array of biological or chemical probes bonded to a support by magnetic coupling by means of a fixing vector, the method comprising the following steps:
- A) bonding probes to the fixing vector;
- and
- B) depositing probes coupled to the fixing vector on a magnetic support in an organized array.
19/ A method of manufacturing an organized array of biological or chemical probes bonded to a support by magnetic coupling by means of a fixing vector, the method comprising the following steps:
- A) magnetically coupling a fixing vector at known coordinates on a support;
- and
- B) bonding probes to the fixing vectors.
20/ A method according to claim 10, characterized in that the step of coupling the fixing vector to the support is performed by using a mask.
21/ A method of manufacturing an organized array of biological or chemical probes bonded to a support by magnetic coupling by means of a fixing vector, the method comprising the following steps:
- A) bonding probes to the fixing vector;
- B) depositing probes coupled to a fixing vector on a non-magnetized slide, in an organized array;
- and
- C) transferring the array of step B) onto a magnetic support.
22/ A method according to claim 18 or claim 21, for making an organized array of probes constituted by single-strand nucleic acid molecules, the method comprising the following steps:
- A) amplifying probes from nucleotide primers, wherein at least one of which is biotinylated;
- B) denaturing the probes by heating, followed by fast cooling;
- C) coupling the biotinylated probes on beads that are paramagnetic, superparamagnetic, or ferromagnetic, and bonded to avidin or streptavidin;
- D) magnetically precipitating the probes bonded to the beads;
- E) removing unbonded probes by washing;
- F) resuspending the probes at a selected concentration;
- and
- G) depositing the probes on a support.
23/ A method according to any one of claims 18 to 22, in which the step of bonding the probe to the fixing vector is performed with a concentration of probes that is saturating for the fixing vector.
24/ A support presenting structured permanent magnetization for making an array of biological or chemical probes, using a method according to any one of claims 18 to 23.
25/ A support according to claim 24, in which the structured magnetization is the result of static magnetization decoupled from any electric field and is inert for any charged molecule.
26/ An organized array of fixing vectors coupled to a support by magnetic interaction between the support and the fixing vectors.
27/ A pair produced by coupling a nucleic acid probe and a fixing vector, in which a fixing vector is coupled to a number of nucleic acid molecules lying in the range 103 to 108, and preferably in the range 105 to 107.
28/ The use of a pair produced by coupling a biological or chemical probe and a fixing vector, to make an organized array of biological or chemical probes according to any one of claims 1 to 17.
29/ A kit for making an organized array of biological or chemical probes, the kit comprising a magnetic support and fixing vectors.
30/ A kit according to claim 29, in which the fixing vector carries elements capable of bonding specifically to a biological probe.