EP2250122A1 - Procede de fonctionnalisation de la paroi d'un pore. - Google Patents
Procede de fonctionnalisation de la paroi d'un pore.Info
- Publication number
- EP2250122A1 EP2250122A1 EP09718040A EP09718040A EP2250122A1 EP 2250122 A1 EP2250122 A1 EP 2250122A1 EP 09718040 A EP09718040 A EP 09718040A EP 09718040 A EP09718040 A EP 09718040A EP 2250122 A1 EP2250122 A1 EP 2250122A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- pore
- activatable
- electro
- entities
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
- B81C1/00015—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems
- B81C1/00206—Processes for functionalising a surface, e.g. provide the surface with specific mechanical, chemical or biological properties
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means
- G01N33/48721—Investigating individual macromolecules, e.g. by translocation through nanopores
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81B—MICROSTRUCTURAL DEVICES OR SYSTEMS, e.g. MICROMECHANICAL DEVICES
- B81B2203/00—Basic microelectromechanical structures
- B81B2203/03—Static structures
- B81B2203/0353—Holes
Definitions
- the present invention relates to a method of functionalization and more particularly to bio-functionalization of at least a portion of the wall of a pore.
- pore or channel or capillary denotes any cavity emerging from a material or not.
- Silanization firstly, performs the covalent grafting of organosilanes to the surface of materials such as glass or silicon. This process most often consists in first carrying out functionalization with a reactive group which will then allow the immobilization of the molecule of interest (Iqbal, S. et al., "Solid state nanopore channels with DNA selectivity", Nature Nanotechnology , 2007, 2: 243 and following, Karnik et al., Nano-Letters, 2007, 7 (3): 547 et seq .; Kim, Y.-R. et al., Biosensors & Bioelectronics, 2007. 22: 2926 and following, Wanunu, M.
- a recent article by Joakim Nilsson et al., Entitled “Localized Functionalization of Single Nanopores” discloses the use of a focused nano-beam or nanoFIB (FIB: " Focused Ion Beam ”) for the creation of a pore in a silicon nitride surface.
- FIB Focused Ion Beam
- this process is multi-step and requires prior silanization of the support.
- the application of these structures is most often connected to the connectors, which leads most authors, if necessary, to dissolve the matrix after creation of the polymer tubes.
- the pore is here only a "mold", creator of the cylindrical form of the generated polymers, and is not intended to be used as an active support.
- two different processes are known and used: chemical polymerization and electropolymerization.
- Patent Applications FR 2 787 582 and FR 2 784 466 relate to a conventional technique of electropolymerization in which an electrode is disposed at the bottom of a non-emergent frustoconical microcuvette and another electrode in an electrolyte, in an unspecified position. .
- this known technique makes it possible to deposit only one of the electrodes.
- the invention relates to a method for performing a functionalization of a pore located on its surface, while simplifying the process.
- the basic idea of the invention is to generate in the pore a voltage gradient of electrical apt to allow a deposit on the walls of the pore.
- the invention thus relates to a process for functionalizing at least part of the wall of at least one pore of a support material, characterized in that it comprises: a) bringing the pore into contact with a solution of entities electro-activatable and position two electrodes in said solution, on either side of the pore, so as to create inside the pore and when an electrical signal is applied between the two electrodes, a voltage drop, in particular greater than 1000 V / m, capable of generating a deposit located on said wall. b) applying an electrical signal, potential difference or current, between the two electrodes to perform said functionalization.
- an electrode is disposed at the bottom of a non-emergent cavity or at the bottom of the pore.
- the field can reach 10 6 V / m or more.
- the electrical signal can be constant or modulated as a function of time (periodic or not, pulsed, modulated in amplitude or in frequency, in staircase, in ramp, etc.).
- the support is not necessarily conductive. There is no need to line the inside of the pores with a conductive layer as described for electropolymerization, which greatly simplifies the experimental procedure.
- the support consisting of organic or inorganic material, may be of insulating, semiconductive or conductive nature.
- Remote electropolymerization does not require the presence of an oxidizing chemical agent.
- the remote electropolymerization process is feasible in a single handling step.
- the preferential formation of the polymer on all or part of the wall of the pore can be explained by the fact that, an electric signal being applied on either side of the pore, the voltage drop that it produces is mainly localized to the pore. interior of the pore resulting in a strong potential gradient inducing a preferential formation of the polymer.
- the method may include at least one iteration of a and b with a second solution of electro-activatable entities.
- These entities may be the same or advantageously different entities, which allows in particular to have layers deposited on each other or side by side.
- the pore is open at both ends and a solution is placed in two compartments in each of which opens an end of the pore, at least one of the two compartments containing said electro-activatable entities.
- the pore has a single emergent end and one of the two electrodes is placed at the bottom of the pore, the other electrode being placed in a compartment in communication with the open end of the pore. After b, it can be provided a rinse.
- the support material may be silicon-based.
- the electro-activatable entities may be electropolymerizable monomers, especially pi-conjugated conducting monomers, preferably a pyrrole, or else carry electrograftable functions, in particular diazonium groups, or else be chosen from metals, metal oxides, catalytic particles, salts and metal complexes or consist of an electrophoretic paint.
- the electro-activatable entity solution may include ligands.
- the electro-activatable entity solution may comprise a mixture of electro-activatable entities, in particular an electropolymerizable monomer and said electro-activatable entities coupled to ligands, for example grafted with an oligonucleotide.
- the solution may have an oligonucleotide probe (pyrrole-oligonucleotide), or more generally pyrrole coupled to a biomolecule.
- the solution of electro-activatable entities may include doping ions of interest, in particular heparin and / or chondroitin.
- the support material may be based on silicon.
- the electrical signal may be a voltage of between 10 mV and 500 V and preferably between 100 mV and 10 V.
- the criterion to be respected is that the field inside the pore is sufficient to generate a deposit on its wall.
- the voltage difference can be applied for a duration of between 10 ⁇ s and 100 s and more particularly between 10 ms and 100 s, for example in the form of a pulse. The time of application of the voltage determines the thickness of the deposit.
- the concentration of electro-activatable entities can be broadly ranging from 1 nM to 500 mM.
- the method may have a step of detaching the electro-activatable entities from the support, for example by destroying this last or under the action of ultrasound.
- the support may have at least one flared functionalization region (with or without bearings) which extends the wall of a pore.
- the electro-activatable entities may comprise probe molecules, and the method may comprise a step of association by recognition in particular of hybridization with complementary target molecules.
- the method may then include a step of denaturing said association by recognition, possibly followed by a step of new association by recognition, including rehybridization.
- the method thus allows the affinity association of an entity of interest and allows the manufacture of molecular assemblies.
- FIG. 1 is a block diagram illustrating the method according to the invention
- FIGS. 2a to 2c represent a cell intended to receive a chip having a pore (assembly a), FIGS. 3 and 4 illustrate a mounting adopted on a multi-pores chip, FIG. 4 being a detail relating to the pore Pi. in the conditions of experience,
- FIG. 5 illustrates the format of the fluorescence test used to validate the functionalization of the pores
- FIGS. 6, 7, 8a and 8b represent different pore profiles used in the examples.
- FIGS. 9a and 9b show two examples of non-emerging pore profiles.
- the present invention relates to a process for functionalizing the surface of a pore by an organic or inorganic entity, in particular by a polymer, which has been generated electrically by the application of an electrical signal, in particular a difference of electrical potential on both sides of the pore. It makes it possible to carry out functionalization of pores or channels regardless of their size (for example of diameter between 1 nm and 5 mm), in particular pores or channels of micrometric and / or nanometric size, by: active groups allowing low energy interactions, for example:
- molecular or biomolecular recognition groups such as, for example, biomolecules, reactive chemical groups or ion chelators.
- organic or inorganic entities in particular for the purpose of reducing the pore opening diameter.
- pore or "channel” or “capillary” means any cavity, open or not, located in a material. Their spatial distribution on the support can be defined (case of a machined membrane for example) or statistical (typical case of a frit). The invention relates to any pore size.
- a pore 1 (FIG. 1) is placed between two sealed compartments 3 and 4 containing a solution 2 of electro-activatable entities which also bathes the pore 1.
- a potential difference for example 2 V, is applied by a voltage source 5 between two electrodes. 6 and 7 disposed on either side of the pore 1, at a distance of a few millimeters from one another.
- the electro-activatable entity may be chosen in particular from:
- Electropolymerizable monomers such as pyrroles, thiophenes, indoles, anilines, azines, phenylenevinylenes, phenylenes, pyrenes, furans, selenophenes, pyrridazines, carbazoles, acrylates, methacrylates and their derivatives.
- the electropolymerizable pattern is a pyrrole.
- This monomer is easily functionalizable by an entity of interest.
- polypyrrole is a biocompatible polymer, stable in air and in solution at physiological pH, which is an asset in the context of an application in the field of biosensors. Derivatives carrying electrograftable functions such as groups diazonium
- Metals and metal oxides for example iridium oxide, catalytic particles, salts and metal complexes
- the porous support may be of organic and / or inorganic nature and indifferently conductive, semiconducting or insulating electric. Semiconductor materials such as silicon or its oxide and nitride derivatives are preferably used.
- Example I the monomer used is pyrrole.
- polypyrrole is a polymer which has the advantage of being biocompatible and is therefore very interesting for developing biosensors. It also has the advantage of being stable in the handling conditions of biochemical tests (physiological pH, aqueous buffers, presence of oxygen, etc.). It is also a conductive polymer that has a hydrophilic character allowing its use in biological systems. Moreover, the synthesis of pyrrole-biomolecule conjugates is very well controlled and is done with a good yield.
- Polypyrrole, polycarbazole, polyaniline, PEDOT 1 , polyindole, and polythiophene belong to the group of pi-conjugated conductive polymers. It is known that the corresponding monomers are electropolymerizable, that is to say that they lead to the formation of a polymer under the effect of the application of anodic potential on the surface of an electrode. These entities therefore behave in the same way taking into account solvent and oxidation conditions that are not identical from one entity to another.
- the pyrrole is aliquoted at a concentration of 1 M in acetonitrile solvent and then stored at -20 ° C.
- the pyrrole bearing an oligonucleotide was prepared according to the protocol described in Patent Application FR 2 703 359.
- the DNA sequences used are the following: Py-probeZip6: Py 5 ' - (T) 10 - GAC CGG TAT GCG ACC TGG TAT GCG 3 ' (Py-SEQ ID NO.1)
- Target-Zip6-bio biotin- 5 ' CGC ATA CCA GGT CGC ATA CCG GTC 3' (biotin-SEQ ID NO: 2)
- the chips used are membranes of silicon oxide or silicon nitride. They comprise nine pores of micrometric size distributed over a surface of 2x2 cm 2 .
- Buffers used (given as an indication):
- Hybridization Buffer 0.02M Na 2 HPO 4 ZNaH 2 PO 41 I 1 IM NaCl 1 5.4mM KCl, 4% v / v 5OX Denhardt, 0.2% v / v Salmon sperm DNA, 0 , 3% v / v Tween 20 at pH 7.4
- Rinse buffer PBS 5 tablets / L, NaCl 23,375 g / L, Tween 20 0.15% v / v.
- Electropolvmerisation cell see FIGS. 2a to 2c:
- the material of this cell C is "DeIMn" (registered trademark) polyoxymethylene said “DeIMn POM”.
- the cell is split into two sealed compartments 3 and 4 by introducing into its rectangular receptacle 34 a support part 8 comprising one or more pores 1.
- Platinum wires for example can be introduced into each of the compartments 3 and 4 through the openings 9i. and 9 2 of the cover 9 to form the electrodes.
- 9 2 and 9 4 denote the cover mounting openings 9 of the cell C, and 5 and 9 9 6 screw mounting holes on the cell C.
- mounting chip "multipore"
- the chip 10 has 9 pores P 1 Pg. Each pore is isolated between two watertight compartments (3i, 4i, 3 2 , 4 2 , .... 3 9l 4 9 ). In other words, it is possible to put each of the nine pores Pi, P 2 Pg which have the same diameter or not in contact with a different solution Si, S 2 S 9 of electro-activatable entities.
- Each of these compartments is equipped with electrodes to which is applied a given electrical potential difference and which are arranged at a distance d from the end of each pore.
- the distance d may or may not be the same for the two electrodes of the same pore. It can be different from one pore to another depending on the requirements of functionalization.
- the solutions Si, S 2 , Sg may or may not be the same.
- a PT "multi-channel" potentiostat makes it possible to apply these voltages simultaneously (even if the values are different).
- Each pore (P 1 , ... P 9 ) of a chip 10 is positioned between two sealed compartments 3i, 4i, ...; 3g, 4 9 which here have a volume of 10 .mu.l.
- the assembly comprises one or two printed circuits 21, 22 (FIG 4) having an integrated circular electrode 23, 28 connectable to an external potentiostat.
- In at least one of the electrodes are drilled two holes 26, 27 for introducing and discharging liquid via polytetrafluoroethylene capillaries.
- Two sealed compartments 3 1 and 4 1 are created between the chip 10 and the printed circuits 21 and 22 by means of O-rings 24 and 25 (FIG. 4).
- the electrodes integrated in the printed circuits or electrodes (not shown metal wires) introduced into capillaries on either side of the pore, or electrodes dipping directly into a compartment.
- a plastic card (not shown) is used in place of the printed circuit.
- the chip is cleaned (67% sulfuric acid, 33% hydrogen peroxide v / v) in a clean room to remove any organic contaminants.
- the chip is soaked for 10 min in the solution and then rinsed through a water circulation until a resistivity of 9 M ⁇ .m.
- the chip is then dried in an oven at 180 0 C for 10 min. It can then be stored at room temperature.
- a polymerization solution containing 20 mM pyrrole and 5 ⁇ M py-Zip6 probe in electropolymerization buffer was used to make polymer deposits in the pores.
- the chip comprising through pores is introduced so as to be between two compartments.
- the polymerization solution is introduced into the two compartments.
- An electrode is inserted in each compartment and a potential difference equal to 2 V is applied.
- a voltage of between 10 mV and 500 V and preferably between 100 mV and 10V can be used depending on the pore size, the aim being to obtain a sufficiently high field inside. pore so that the deposit takes place on its wall.
- the tracking of the deposition process is done by drawing the curve of the evolution of the intensity of the current as a function of time: the shape of this curve (presence or absence of an electrical signal) makes it possible to see if the liquid has penetrated to inside the pore (electrical contact) or not (no electrical signal).
- the chip is then removed from the assembly and rinsed with water, dried with compressed air and kept dry at 4 ° C. d) Verification of functionalisation by fluorescence microscopy
- fluorescence microscopy is used.
- the format of the test used is shown in Figure 5. It is performed by depositing drops of 15 .mu.l of liquid on a pore.
- the pore is first saturated with Hybridization Buffer (5 min at room temperature). Then a drop of biotinylated target at 100 nM hybridization buffer is added (15 min, room temperature). The chip is then thoroughly rinsed with the Rinse Buffer. Each pore is then incubated in a 10% streptavidin-phycoerythrin (SAPE) solution
- the first is connected to the counter electrode coupled to the reference electrode of the potentiostat and the second to the working electrode.
- a potential of 2 V is applied for a given time (between 100 ms and 1 s) between the two electrodes (working electrode and counter-electrode).
- the chip is then removed from the cell, thoroughly rinsed with water and then dried with compressed air and stored at 4 ° C.
- R f 35: pore 70 ⁇ m in diameter in a membrane of 2 microns thick and 500 microns side (see Figure 6).
- a fluorescence emission is observed in the form of a luminous circle present on each side of the chip. Its dimensions correspond to those of the pore contour, which allows to deduce that the functionalization technique is effective and allows a polymer deposit located on the walls of a pore of micrometer size.
- Scanning electron micrographs show a thin deposition layer - on the order of 30 nm - on the contour of a pore. This deposit is absent from an unfunctional pore.
- R f 1 with a circular pore 18 ⁇ m in diameter in a membrane 20 ⁇ m thick: case of a pore pierced in a square membrane of 50 ⁇ m side and 20 ⁇ m thick.
- the deposition is carried out at a voltage of 2 V applied for 100 milliseconds (FIG. 7).
- R f 0.25 pore diameter 2 microns in a membrane of 8 microns thick.
- the pore with a size of 2 ⁇ m is at the bottom of a cone with a larger diameter equal to 10 ⁇ m.
- the environment of the pore is said to be of the "funnel" type (FIGS. 8a and 8b).
- Fluorescence microscopy photographs show that the pore wall has been functionalized in a localized manner, as well as the contour of the top of the cone (10 ⁇ m in size).
- a multi-pores chip having pores of variable form R f form and O 2 plasma-processed is placed within the assembly b described above ( Figures 3 and 4).
- the polymerization solution is introduced successively into the two compartments. It consists of 20 mM pyrrole and 5 ⁇ M pyrrole-probe-Zip ⁇ in electropolymerization buffer. Then, two electrodes are positioned on each side of the chip. The first is connected to the auxiliary and reference electrodes of the potentiostat and the second to the working electrode. A potential of 2V is applied for 100 ms between the two electrodes. The chip is then removed from the cell, thoroughly rinsed with water and then dried with compressed air and stored at 4 ° C. ii) Results
- iridium oxalate solution is prepared according to the following protocol (described in the article by A. M. Marsouk, Analytical Chemistry,
- the iridium oxalate solution is introduced successively into the two compartments. Then, two electrodes are positioned on each side of the chip. The first is connected to the auxiliary and reference electrodes of the potentiostat and the other to the working electrode. A potential of
- 0.80 V or 0.85 V or 0.90 V is applied for a duration of 5 s or 10 s.
- the monitoring of the deposit is done by chronoamperometry to check the good electrical contact through the pore.
- EXAMPLE IV OBTAINING STRUCTURED OBJECTS BY REMOTE ELECTRQDEPT TECHNOLOGY: i) Creation of a polypyrrole deposit: A polycarbonate membrane, comprising pores of nanometric size, can be inserted indifferently in assemblies a or b. Deposits of pyrrole / pyrrole-coupled copolymer with an oligonucleotide can then be obtained inside these pores following the previously described protocol.
- FIGS. 9a and 9b are two variants of non-emerging pore shapes with an electrode 61 which covers all or part of the bottom of the cavity 60.
- the zone of the pore where the deposit takes place corresponds at the narrow region 62, 62 'which concentrates the field.
- the shape of the pore makes it possible to specifically locate the deposit on only a part of its wall.
- the polarization of the electrode 61 at the bottom of the cavity 60 may be anodic or cathodic to respectively form a deposit of the same polymer on the surface of the electrode 61, in addition to the deposit on the necking region 62, 62 '. CONCLUSION:
- the technical method of functionalization according to the invention is effective and relatively easy to put into practice.
- This new technique makes it possible to effectively control the location of the functionalization of the surface of a pore by reactive groups. Indeed, the latter is essentially related to the organization of electric field lines within the pore, itself dependent on the structure of the pore environment (ie its geometry).
- the method according to the invention has the advantage of being inexpensive: in financial terms because only reduced equipment is needed: potentiostats, electrodes ... in terms of time because the filing procedure only takes a few minutes.
- the experimental device is more compact and easily transportable.
- the strategy is adaptable to any type of porous support, organic or inorganic, conductive, semiconductor or insulating, regardless of the pore size.
- the chronoamperogram measured during the deposition for example of polypyrrole
- the chronoamperogram measured during the deposition has a non-intensity signal. no.
- Electro-activatable entities in particular electropolymerizable (pyrroles, thiophenes ...), which can be functionalized, this technique is perfectly transferable to the immobilization within pores of active groups involved in low energy interactions such as ionic groups, peptides , antibodies, enzymes, ion chelators for example.
- the polymer may also serve as a "starting layer" for localized deposition, in particular by using doping anions of interest, such as polysaccharides (heparin for example promoting cell adhesion (Zhou et al., Reactive & Functional Polymers, 1999). 39: 19 and following)) or surfactants.
- doping anions of interest such as polysaccharides (heparin for example promoting cell adhesion (Zhou et al., Reactive & Functional Polymers, 1999). 39: 19 and following)
- surfactants such as polysaccharides (heparin for example promoting cell adhesion (Zhou et al., Reactive & Functional Polymers, 1999). 39: 19 and following)
- Multi-layer type stacks can be envisaged from the deposits obtained by "remote electrodeposit". It is thus possible to prepare a localized deposit (first layer) having for example a certain surface charge or a reactive chemical group which favors the fixing of a second layer of organic or inorganic entities given relative to the blank support.
- the technique has experimentally allowed the implementation of a DNA immobilization, the latter being a biomolecule having a modular aspect, that is to say that can be used as a biomolecular recognition element for immobilization via hybridization of d a molecule of interest functionalized by complementary DNA targets.
- the method has also made it possible to immobilize an entity of biological interest, biotin, by hybridization with immobilized DNA probes with a biotinylated complementary target, which underlines the modular aspect of the technique.
- the experimental device can further integrate thermal and optical devices allowing for example crosslinking experiments or a visualization of the organization of deposits made.
- Biofunctionalized porous membranes can find applications in the field of health, in particular for the detection of (bio) molecules present in small quantities in biological samples. Many research teams have thus oriented their work towards the design of systems for the detection of single molecules. These molecular Coulter counters have provided encouraging results with proteinaceous pores (Vercoutere, W. et al., Nature Biotechnology, 2001. 19: 248, Bayley and Cremer, Nature, 2001. 413: 226 et seq.).
- the method is also suitable for applications in the field of micro- or nano-chromatography (ion exchange, steric exclusion, affinity or adsorption chromatography) for the purification of (bio) molecules.
- micro- or nano-chromatography ion exchange, steric exclusion, affinity or adsorption chromatography
- Functionalized pores may also be useful for capturing bacteria or cells, including immobilizing heparin or chondroitin within a pore.
- Functionalised porous membranes also typically find applications in purification and filtration systems (water, effluents, etc.), the presence of chelators or ion exchangers on the surface of the pores being capable of selectively separating certain components of the liquid passing through the membrane.
- the immobilization of catalytic particles, for example containing metals such as palladium or platinum in pores, via the process described, could allow the creation of micro- or nano-reactors for carrying out chemical reactions such as hydrogenations. for example.
- the fact of being able to parallelize these reactions by using pore networks distributed in a membrane makes it possible to make combinatorial micro / nano-chemistry. Electrodeposition of metals by this technique can also find applications in the fields of catalytic converters (gas phase catalysis).
- the method is finally compatible with the implementation of molecular imprinting techniques, which opens up interesting applications in the field of capillary electrophoresis, for example.
- electro-activatable entities for which the possible presence of DNA probes has been mentioned above, may more generally optionally include ligands, namely: molecular and / or biomolecular recognition elements including nucleotides, oligonucleotides, polynucleotides, DNA, RNA, PNA, peptides, polypeptides, antibodies, antigens, enzymes, proteins, amino acids, glycopeptides, biotins, haptens, sugars, oligosaccharides, polysaccharides , lipids, glycolipids, steroids, hormones, receptors.
- ligands namely: molecular and / or biomolecular recognition elements including nucleotides, oligonucleotides, polynucleotides, DNA, RNA, PNA, peptides, polypeptides, antibodies, antigens, enzymes, proteins, amino acids, glycopeptides, biotins, haptens, sugars, oligosacchari
- affinity groups in particular ion chelators and ion exchangers.
- the particles may be and / or contain biological cells and / or components and / or cellular products, including cell lines and / or globules and / or liposomes and / or cell nuclei and / or chromosomes and / or or strands of DNA or RNA and / or nucleotides and / or ribosomes and / or enzymes and / or antibodies and / or proteins and / or proteins and / or peptides and / or principles active agents and / or parasites and / or bacteria and / or viruses and / or pollens and / or polymers and / or biological factors and / or stimulants and / or growth inhibitors and / or suspension in a liquid, and / or bioparticles suspended in a solution, and / or molecules.
- biological cells and / or components and / or cellular products including cell lines and / or globules and / or liposomes and / or cell nuclei and /
- the manipulated particles may be and / or contain insoluble solid particles such as magnetic particles and / or dielectric particles, or conductive particles, or functionalized particles, or pigments, or dyes, or protein crystals, or powders, or polymer structures, or insoluble pharmaceutical substances, or carbon fibers, or yarns, or nanotubes, or small aggregates (clusters) formed by agglomeration of colloids of o-groups having surface features • in terms of pH, especially weak acid / base pairs and amphoteric compounds and / or in terms of hydrophilicity and / or hydrophobicity and / or amphiphilism, and / or in terms of polarity,
- insoluble solid particles such as magnetic particles and / or dielectric particles, or conductive particles, or functionalized particles, or pigments, or dyes, or protein crystals, or powders, or polymer structures, or insoluble pharmaceutical substances, or carbon fibers, or yarns, or nanotubes, or small aggregates (clusters) formed by agglomeration of colloids
- the functionalization layer deposited on the walls of the pore comprises, for example, interaction or reaction means with molecules and / or biomolecules.
- These include modular groups such as DNA, photoactivatable such as benzophenone, electroactivables such as the electro-activatable entities mentioned above, or even thermoactivables such as thermosetting polymers.
- the ligand must be coupled to an electro-activatable entity to enable functionalization by the method of the present invention. It is not necessary that there be in the solution both electro-activatable entities and electro-activatable entities coupled to a ligand: there can also be only electro-activatable entities coupled to a ligand, or else only electro-activatable entities.
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Abstract
Description
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Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR0800601A FR2927169B1 (fr) | 2008-02-05 | 2008-02-05 | Procede de fonctionnalisation de la surface d'un pore |
PCT/FR2009/000133 WO2009109727A1 (fr) | 2008-02-05 | 2009-02-05 | Procede de fonctionnalisation de la paroi d'un pore. |
Publications (1)
Publication Number | Publication Date |
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EP2250122A1 true EP2250122A1 (fr) | 2010-11-17 |
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
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EP09718040A Withdrawn EP2250122A1 (fr) | 2008-02-05 | 2009-02-05 | Procede de fonctionnalisation de la paroi d'un pore. |
Country Status (6)
Country | Link |
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US (1) | US20110174629A1 (fr) |
EP (1) | EP2250122A1 (fr) |
JP (1) | JP2011511167A (fr) |
CA (1) | CA2714185A1 (fr) |
FR (1) | FR2927169B1 (fr) |
WO (1) | WO2009109727A1 (fr) |
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FR2950044B1 (fr) * | 2009-09-11 | 2011-12-09 | Commissariat Energie Atomique | Procede de preparation d'une surface structuree fonctionnelle et surface obtenue par le procede |
US9422154B2 (en) | 2010-11-02 | 2016-08-23 | International Business Machines Corporation | Feedback control of dimensions in nanopore and nanofluidic devices |
US8852407B2 (en) * | 2011-01-28 | 2014-10-07 | International Business Machines Corporation | Electron beam sculpting of tunneling junction for nanopore DNA sequencing |
FR2975986A1 (fr) * | 2011-06-01 | 2012-12-07 | Commissariat Energie Atomique | Procede de fonctionnalisation de la paroi d'un pore. |
WO2012173592A1 (fr) | 2011-06-13 | 2012-12-20 | Empire Technology Development Llc | Système de membranes programmables |
JP5795686B2 (ja) * | 2011-06-13 | 2015-10-14 | エンパイア テクノロジー ディベロップメント エルエルシー | 多孔膜上への機能性および再使用可能電着コーティング |
US9738984B2 (en) | 2011-06-13 | 2017-08-22 | Empire Technology Development Llc | Reliable point of use membrane modification |
US10029915B2 (en) | 2012-04-04 | 2018-07-24 | International Business Machines Corporation | Functionally switchable self-assembled coating compound for controlling translocation of molecule through nanopores |
US9777389B2 (en) * | 2012-05-07 | 2017-10-03 | The University Of Ottawa | Fabrication of nanopores using high electric fields |
US9057693B2 (en) | 2013-04-29 | 2015-06-16 | International Business Machines Corporation | Silicon oxide nanopore wetting and stabilization by molecular coating |
US9714933B2 (en) * | 2014-01-28 | 2017-07-25 | International Business Machines Corporation | Micro-droplet fluidic cell for fast ionic current detection using nanopores |
DE102014111984B3 (de) * | 2014-08-21 | 2016-01-21 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Fluidische Gigaohm-Dichtung für Transmembranproteinmessungen |
US11333626B2 (en) * | 2016-02-22 | 2022-05-17 | Hitachi, Ltd. | Biological sample analysis chip, biological sample analyzer and biological sample analysis method |
US10641726B2 (en) | 2017-02-01 | 2020-05-05 | Seagate Technology Llc | Fabrication of a nanochannel for DNA sequencing using electrical plating to achieve tunneling electrode gap |
US10889857B2 (en) | 2017-02-01 | 2021-01-12 | Seagate Technology Llc | Method to fabricate a nanochannel for DNA sequencing based on narrow trench patterning process |
US10640827B2 (en) | 2017-02-01 | 2020-05-05 | Seagate Technology Llc | Fabrication of wedge shaped electrode for enhanced DNA sequencing using tunneling current |
US10731210B2 (en) | 2017-02-01 | 2020-08-04 | Seagate Technology Llc | Fabrication of nanochannel with integrated electrodes for DNA sequencing using tunneling current |
US10761058B2 (en) | 2017-02-01 | 2020-09-01 | Seagate Technology Llc | Nanostructures to control DNA strand orientation and position location for transverse DNA sequencing |
US10752947B2 (en) | 2017-03-09 | 2020-08-25 | Seagate Technology Llc | Method to amplify transverse tunneling current discrimination of DNA nucleotides via nucleotide site specific attachment of dye-peptide |
US20180259475A1 (en) | 2017-03-09 | 2018-09-13 | Seagate Technology Llc | Vertical nanopore coupled with a pair of transverse electrodes having a uniform ultrasmall nanogap for dna sequencing |
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Publication number | Priority date | Publication date | Assignee | Title |
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US4204918A (en) * | 1978-09-05 | 1980-05-27 | The Dow Chemical Company | Electroplating procedure |
US6095148A (en) * | 1995-11-03 | 2000-08-01 | Children's Medical Center Corporation | Neuronal stimulation using electrically conducting polymers |
US5753316A (en) * | 1997-01-14 | 1998-05-19 | Ppg Industries, Inc. | Treatment of metal parts to provide improved sealcoat coatings |
US20020083586A1 (en) * | 1998-04-10 | 2002-07-04 | Takahiro Iijima | Process for producing multilayer circuit board |
FR2784466B1 (fr) * | 1998-10-13 | 2002-10-18 | Commissariat Energie Atomique | Micro-systeme a multiple points d'analyse chimique ou biologique mettant en oeuvre un couplage entre les sondes et un substrat |
FR2787582B1 (fr) * | 1998-12-16 | 2001-01-12 | Commissariat Energie Atomique | Procede de fabrication d'une biopuce et biopuce |
ATE317711T1 (de) * | 1999-12-03 | 2006-03-15 | Yissum Res Dev Co | Elektropolymerisierbare monomere und polymerbeschichtungen auf implantierbaren geräten |
FR2843828A1 (fr) * | 2002-08-26 | 2004-02-27 | Commissariat Energie Atomique | Support de garniture et procede de garniture selective de plages conductrices d'un tel support |
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2008
- 2008-02-05 FR FR0800601A patent/FR2927169B1/fr not_active Expired - Fee Related
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2009
- 2009-02-05 US US12/866,340 patent/US20110174629A1/en not_active Abandoned
- 2009-02-05 JP JP2010545525A patent/JP2011511167A/ja active Pending
- 2009-02-05 EP EP09718040A patent/EP2250122A1/fr not_active Withdrawn
- 2009-02-05 WO PCT/FR2009/000133 patent/WO2009109727A1/fr active Application Filing
- 2009-02-05 CA CA2714185A patent/CA2714185A1/fr not_active Abandoned
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US20110174629A1 (en) | 2011-07-21 |
JP2011511167A (ja) | 2011-04-07 |
FR2927169B1 (fr) | 2013-01-11 |
FR2927169A1 (fr) | 2009-08-07 |
WO2009109727A1 (fr) | 2009-09-11 |
CA2714185A1 (fr) | 2009-09-11 |
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