EP1032826A1 - Procede de production des structures d'organisation laterale sur des surfaces support - Google Patents

Procede de production des structures d'organisation laterale sur des surfaces support

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Publication number
EP1032826A1
EP1032826A1 EP98966157A EP98966157A EP1032826A1 EP 1032826 A1 EP1032826 A1 EP 1032826A1 EP 98966157 A EP98966157 A EP 98966157A EP 98966157 A EP98966157 A EP 98966157A EP 1032826 A1 EP1032826 A1 EP 1032826A1
Authority
EP
European Patent Office
Prior art keywords
molecules
electrode
electrodes
immobilized
group
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Withdrawn
Application number
EP98966157A
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German (de)
English (en)
Inventor
Vladimir M. Mirsky
Michael Riepl
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wolfbeis Otto Samuel
Original Assignee
Wolfbeis Otto Samuel
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Filing date
Publication date
Application filed by Wolfbeis Otto Samuel filed Critical Wolfbeis Otto Samuel
Publication of EP1032826A1 publication Critical patent/EP1032826A1/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • G01N33/5438—Electrodes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001—Enzyme electrodes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00653—Making arrays on substantially continuous surfaces the compounds being bound to electrodes embedded in or on the solid supports
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00659—Two-dimensional arrays

Definitions

  • the present invention relates to an arrangement on the carrier surface of which molecular layers are immobilized in an electrically addressable manner, a method for electrically addressable immobilization of molecules, a device for carrying out this method and the use of this arrangement as a chemical and / or biosensor, in particular as a multi-sensor system for chemical , biological and physical determinations and for applications in combinatorial synthesis at the interface.
  • Both chemical sensors and biosensors i.e. Comparatively small measuring arrangements are increasingly required to carry out chemical or biochemical analyzes quickly and at the scene. They are of particular advantage to immunological detection systems because they are able to record, for example continuously, concentrations of (bio) chemical substances in a short time and oline complex sample preparation, preferably continuously.
  • a biosensor should therefore be small, which gives another advantage of the application due to the proximity to the location of the examination, e.g. diabetics can determine their blood sugar level in a few minutes with the help of enzymatic biosensors.
  • the immobilization methods known above have many disadvantages which limit the applicability of the methods.
  • the resolution of the micromechanical immobilization limited by the size of the individual spray particles.
  • an optical resolution of approx. 100 ⁇ m is achieved in the best case.
  • protective groups must always be used which require compliance with certain conditions (for example solvents, darkening due to photosensitivity and the like) and which have to be split off again at a certain point in time.
  • the Chrisey et al. (1996) is very time-consuming and costly since a large number of photomasks have to be used.
  • One solution to this problem is to provide an arrangement with electrically addressable immobilized molecules which has a support, one and / or more (1 to n) electrically conductive support surface (s) which is / are arranged on this support and one and / or several immobilized identical and / or different receptor (s).
  • the electrodes are separately assigned either an adsorption potential or a desorption potential, the uncoated electrode locations being kept inert by the desorption potential and the adsorption potential still applied to the already coated electrode locations preventing the desorption of molecules as well as the further adsorption of differently structured molecules, whereby the electrode site is occupied by the first type of molecule and the molecules are immobilized and firmly attached to the support surface and
  • the process is complete when a constant signal value is reached. e.g. Capacitance, impedance, or resonance frequency value.
  • the advantage of the inventive method is that the potential can be controlled within and outside the potential stability range, the desorption or adsorption of molecules with the ⁇ nde ⁇ * ing. With this principle it is possible to build up a certain lateral structure of the chemically adsorbed layer.
  • This method can be carried out with the device according to the invention, which consists of a changing device (1) for sample containers.
  • a pump (2) a flow cell (3) into which the desired molecule or receptor or into which the desired molecules or receptors are transported by means of the pump (2), a support (5) on which n electrodes are arranged, the support (5 ) with the n electrodes in the flow-through cell (3), a multiplexer (6), an address bus (7) which controls the multiplexer (6), whereby to the electrode (s) separately either an adsorption potential (8) or a desorption potential (9).
  • a changing device (1) for sample containers.
  • a pump (2) a flow cell (3) into which the desired molecule or receptor or into which the desired molecules or receptors are transported by means of the pump (2)
  • a support (5) on which n electrodes are arranged the support (5 ) with the n electrodes in the flow-through cell (3)
  • a multiplexer (6) an address bus (7) which controls the multiplexer (6), whereby to the
  • Reference electrode (10) is / are applied, and a collecting vessel (4), in which after the immobilization of the molecules by means of the
  • the arrangement according to the invention is used as a chemical and / or biosensor. used in particular as a multi-sensor system.
  • the arrangements according to the invention with the electrically addressable immobilized molecules can also be used for combinatorial synthesis by specifically releasing molecules bound on the electrode surface, which may be identical or different depending on the electrode, by applying the desorption potential and thus as a reaction partner during the system step supplies or if the synthesis was carried out on the surface, the products can be split off directly.
  • Electrodes of any size, shape and number are used for the electrically addressable immobilization, which are applied to a carrier made of a dielectric material. In this dielectric material, electrical leads for sensor spots can optionally be used be introduced. Immobilization takes place according to the schemes shown in FIGS. 1 and 2, which are described below. During the immobilization, different receptors (molecules A, B, C, etc.) that contain one or more thiol groups are specifically bound to different electrodes (/, 2, 3, etc.): receptor A on electrode El, receptor B on Electrode E2, etc. To achieve these bonds, the procedure is as follows: A suitable electrode potential (adsorption potential) is applied to electrode El in liquid electrolyte compared to a reference electrode which supports the binding with thiol groups.
  • a suitable electrode potential adsorption potential
  • the other electrodes (2, 3, etc.) are given an electrode potential (desorption potential) with respect to the same reference electrode, which is sufficient. to suppress the binding of the thiol groups to these electrodes. Therefore, the added receptor molecule A is only bound to the electrode El. Then the receptor molecules A in the liquid electrolyte are replaced by a liquid electrolyte solution with receptor molecules B and these molecules are addressed to the subsequent electrode position 2. Due to the bending of the desorption potential, the uncoated electrode positions are kept inert. The adsorption potential that is still applied to the already coated electrode sites prevents the desorption of molecules as well as the further adsorption of differently structured molecules because the electrode site is completely occupied by the first type of molecule. This process is repeated until the sensor arrangement is completely set up. A firm bond is formed between the immobilized molecule and the electrode. It is thus possible to generate a biosensor or a multisensor system (“.A ⁇ ay”) with any molecular structure.
  • one or more molecular layers for example antibodies, DNA strands via functional groups
  • one or more molecular layers can be attached to any number of (0 to n) electrodes on which the receptors are located, can be chemically and / or physically adsorbed and / or immobilized.
  • the adsorption potential is maintained for all coated electrodes and the liquid electrolyte solution of the molecules to be coupled is simultaneously pumped into the flow cell with a coupling reagent.
  • the coupling reagent can also be added first and after the activation of the base layer (s) the molecules to be coupled are added.
  • the coupling reagent can be dispensed with, for example with the avidin-biotin system, Ni-His tag, hydrophobic-hydrophobic interactions of liposomes with unfunctionalized alkane thiol chains.
  • rinsing with liquid electrolyte is carried out in order to remove unused or deactivated coupling reagent and / or non-coupled molecules from the cell.
  • this process can be repeated several times until the desired sensor structure is reached step by step.
  • two or more electrodes of the multi-sensor system can be coated with the aid of the adsorption potential and, if necessary, modified with a subsequent physical or chemical adsorption or coupling with one or more identical or different molecular layers certain electrodes have the desorption potential, after the desorption and washing out of the desorbed molecules another coating of these electrodes can be carried out, according to the method described above, this process can be repeated until the desired multisensor arrangement is produced.
  • Particularly suitable coupling reagents are carbodiimides and their derivatives, and N-succinimides and their derivatives.
  • any solid dielectric substrate in particular silicon, glass, non-conductive plastic such as Teflon, PVC, PE, and a conductive or semiconductive substrate which is insulated from the electrode (s) by a dielectric layer is suitable as a carrier.
  • Thin conductive materials that are firmly bonded to the carrier are used as electrodes.
  • Au, Pd, Pt, Ag, alloys such as Au / Pd, Au / Ag, Ag / Pd, GaAs and the like are particularly suitable, doped semiconductors and any other conductive or semiconducting inorganic or organic material such as TCNQ, TTF.
  • the electrodes generally used in electrochemistry such as Ag / AgCl etc. with and without a salt bridge, are used as reference electrodes.
  • Molecules whose binding to the electrodes can be controlled by the electrode potential are used as receptors which are bound to electrodes by means of the method for electrically addressable immobilization according to the invention.
  • These molecules have at least one thiol group or are coupled to at least one thiol group or have a sulfide or disulfide group. They are selected from the group consisting of HS- (CH 2 ) n -X, where n is a number from 2 to 24 and X is H. OH, SH, CH 3 , COOH, NH 2 , and any other molecular fragment stand,
  • Oligonucleotides DNA, RNA, viruses, bacteriophages, prions, oligosaccharides, natural and artificial receptors, redox-active substances, dyes, acids.
  • Bases, epitopes, antigens or antibodies which have at least one thiol group or are coupled to at least one thiol-containing compound.
  • These receptors can also have sulfide or disulfide groups.
  • the thiol groups can either be present originally in such molecules or they can only be introduced by chemical modification.
  • the S atom of the thiol group in all of the compounds mentioned above can be replaced by a Se atom.
  • the molecules that can additionally be arranged on the receptors are selected from the group consisting of toxins.
  • Hormones hormone receptors, peptides, proteins, enzymes, enzyme substrates, cofactors. Medicines, lectins, sugar. Oligonucleotides, DNA, RNA, viruses, bacteriophages, prions, oligosaccharides, natural and artificial receptors, redox-active substances, dyes, acids, bases, epitopes, antigens or antibodies that have at least one functional group or are capable of using the adsorbed receptor interactions, whereby they can be chemically and / or physically adsorbed and / or immobilized on the electrode (s).
  • An adsorption potential is understood to mean an electrical potential in which the binding of the electrode to the molecule is supported and / or maintained.
  • the adsorption potential is at pH values within a range from 4.0 to 8.0 in the range from 0 mV to +600 mV compared to the Ag / AgCl electrode in 100 mM KCI, preferably approximately +300 mV. This potential range is shifted at alkaline pH.
  • desorption potential means a potential outside the stability range defined above.
  • the desorption potential must be high enough to adsorb molecules used during the immobilization process. to prevent.
  • the desorption potential is at a pH within a range from 4.0 to 8.0 in the range from -300 mV or lower compared to the Ag / AgCl electrode in 100 mM KCI. preferably in the range from -600 mV to -1800 mV, in particular in the range from -800 mV to -1400 mV.
  • aqueous solutions organic electrolytes and mixtures thereof as well as a mixture of aqueous electrolytes and organic solvents or organic electrolytes and organic solvents can be used as electro-solutions.
  • Figure 1 shows schematically the device for performing the method for electrically addressable immobilization.
  • FIG. 2 shows the procedure for the electrically addressable coating of a chemo and / or biosensor, in particular a multisensor system (“arrays”) with n individual electrodes, where n is an integer.
  • FIG. 3 shows the reduction in capacity (as a function of time) of an uncoated gold electrode during the adsorption of 6-mercaptohexanoic acid at an electrode potential of +300 mV compared to an Ag / AgCl electrode in 100 mM KCI.
  • FIG. 4 shows the changes in capacitance of an uncoated gold electrode during several cycles of the adsorption / desorption of octanethiol at an electrode potential of -1400 mV compared to an Ag / AgCl electrode in 100 mM KCI. A final rinse completely removes the adsorbed thiol.
  • FIG. 5 shows the change in capacitance of an uncoated gold electrode, to which the desorption (-1400 mV) and adsorption potential (+300 mV) compared to an Ag / AgCl electrode in 100 mM KCI were applied. The adsorption of 6-mercaptohexanoic acid was monitored at both potentials.
  • FIG. 6 shows the changes in capacity of an arrangement consisting of two gold electrodes, in which the electrically addressable immobilization of 6-mercaptohexanoic acid and octanethiol was carried out at electrode potentials of +300 mV and -1400 mV compared to an Ag / AgCl electrode in 100 mM KCI.
  • FIG. 7 shows the change in capacitance of HSA immunosensors when the antigen is added, each symbol type representing a sensor.
  • the device shown in Figure 1 consists of a changing device (1) for sample containers and a pump (2) which transports the desired molecules or receptors into the flow cell (3).
  • n electrodes are arranged on a carrier (5).
  • Either an adsorption potential (8) or a desorption potential (9) is applied separately to these electrodes by means of a multiplexer (6), which is controlled by an address bus (7), with respect to a reference electrode (10).
  • the excess molecules are pumped into a collecting vessel (4) by means of the pump (2).
  • FIG. 2 shows the individual steps of the electrically addressable immobilization of x molecules on n electrodes in detail.
  • (11) represents the addition of a thiol-containing molecule
  • (12) the adsorption of this molecule
  • (13) the rinsing with electrolyte solution.
  • Molecule A is transported into the cell (3) via the pump (2).
  • the molecule A adsorbs on the electrode El, to which the adsorption potential is applied.
  • chemical adsorption on electrodes E2 to En is prevented by the deodorization potential applied there.
  • the adsorption is monitored by means of capacity measurements.
  • the adsorption of molecule A is complete and the cell can be rinsed with electrolyte solution.
  • the electrode potentials are changed in such a way that the adsorption potential is maintained at El and the desorption potential is retained at electrodes E3 to En.
  • the electrical potential is changed by applying the adsorption potential.
  • the molecule B is transported into the cell from the changing device (1) by means of the pump (2). There molecule B only absorbs on E2, because El is already coated with molecule A and the deodorization potential is applied to the electrodes E3 to En. The cleaning is carried out as described above.
  • the coating of the electrodes E3 to En with the molecules C to X is carried out analogously to the electrodes E1 and E2.
  • FIGS 3 to 5 show the changes in capacitance in the electrically addressable
  • Electrodes labeled with ultrapure water and chloroform An aqueous was used as the electrolyte phosphate-buffered KCl solution (100 mM) of pH 6.7 was used.
  • the figures also show the specific capacitance values.
  • the complete removal of the thiols adsorbed at - 1400 mV during the rinsing shows that these molecules were only physically adsorbed on the gold electrode (FIG. 4).
  • the majority of the thiols adsorbed at +300 mV was chemically adsorbed and therefore stable against the rinsing.
  • Figure 6 shows the result with a two-electrode system.
  • Aqueous phosphate-buffered KCl solution 100 mM of pH 6.7 was used as the electrolyte.
  • the addition of 6-mercaptohexanoic acid (16) and the rinsing (15) were carried out at electrode potentials of +300 mV (adsorption potential) for electrode El and -1400 mV (desorption potential) for electrode E2.
  • the addition of octanthiol (14) was carried out for both electrodes at electrode potentials of +300 mV.
  • the electrically addressable immobilization enables the construction of a multi-sensor system that is used, for example, in clinical diagnostics or chemical-biological analysis, e.g. High-throughput screening, can be used. Through the targeted construction of individual electrodes, an ensemble tailored to the user can be produced.
  • enzyme activities in electrolyte solutions or biological liquids, such as blood, urine and the like can be determined.
  • the activity of phospholipase A 2 can be determined by adding a liposome layer to an unmodified alkanethiol monolayer by hydrophobic-hydrophobic interactions. eg hexadecanethiol or octanethiol binds. If phospholipase A 2 is now present in the cell, the lipids are broken down. This can be demonstrated by increasing the capacity signal.
  • Another multi-sensor system can represent the combination of different affinity sensors. Detection of an excess of human serum albumin (HSA) in the urine, a Signs of microalbuminuria, one can use sensors that are constructed as follows.
  • HSA human serum albumin
  • FIGS. 3 to 7 show that it has been possible to adsorb different molecules on specific electrodes by means of electrically addressable immobilization and / or to desorb and / or immobilize. Furthermore it could be shown that the electrodes can be used as a multi-sensor in a system. It has also been possible to provide a detection system with which substances can be detected both quantitatively and qualitatively.
  • Silicon wafer pieces with a size of 3.20 mm x 10.02 mm and a thickness of 450 microns were in a general sputtering process with a gold electrode size 1.56 mm x 1.56 mm (reactive surface) and a lead 10 ⁇ m wide and 6.65 mm long.
  • the electrode was built up from a titanium and palladium layer (adhesion promoter, each 50 ⁇ m thick) and a covering gold layer (200 nm).
  • a silver-plated wire was soldered onto the upper end of the leads as a contact point for the measuring system.
  • the wafer plates were visually checked for damage using a reflected light microscope. The cleaning was done in several steps. First, the wafers were completely immersed in chloroform for 30 minutes.
  • the wafers were immersed in a 1: 1 (v / v) mixture of chloroform and methanol and treated in an ultrasound bath for 10 minutes. Analogous to the chlorofo ⁇ n used, ethanol (99%) and a mixture of ethanol and methanol can also be used in these cleaning steps.
  • the wafers were dried and immersed in a hot 3: 1 (v / v) mixture of concentrated sulfuric acid and 30% hydrogen peroxide solution for 5 minutes. In the last two steps, care was taken that only the reactive electrode surface and a maximum of 4.50 mm of the feed line were immersed in the mixture. The electrodes were thoroughly rinsed with ultrapure water (Millipore: Mili.Qp ⁇ us -185; 18.2 M ⁇ cm "1 ) and dried. All glass and Teflon devices were carefully cleaned according to the previous description before use.
  • the wafer with the sputtered gold electrode was fastened together with an Ag / AgCl reference electrode (surface area approx. 1 cm 2 ) to a Teflon holder, which served as a cover for the measuring cell (snap cover glass, 40 mm x 19 mm) and an opening for the addition and removal of liquids.
  • the cell was filled with electrolyte (100 mM KCI; pH 6.7; approx. 3 ml) until the reactive surface of the gold electrode and the reference electrode (Ag / AgCl) were completely immersed.
  • a magnetic stirrer ensured uniform mixing.
  • a lock-in amplifier with an integrated sine wave generator a constant sinusoidal signal with a frequency of 20 Hz and an amplitude of 10 mV.
  • the adsorption potential (+300 mV) was maintained and as much 6-mercaptohexanoic acid dissolved in the electrolyte was added that the concentration in the measuring cell was 50 ⁇ mol / 1.
  • the adsorption started immediately and was complete after approx. 2.5 hours (compare FIG. 3).
  • the half-life of the coating was approximately 10 minutes and the absolute capacitance value was 4.3 ⁇ F / cm 2, comparable to that of a gold electrode coated in chloroform or ethanol (1 mM 6-mercaptohexanoic acid).
  • the measuring cell was opened, the gold electrodes were rinsed with ultrapure water and immersed in chloroform for about 5 seconds in order to remove physically adsorbed thiol from the electrodes.
  • the reference electrode, measuring cell and stirrer were thoroughly cleaned with ultrapure water, chloroform, ethanol and acetone to remove 6-mercaptohexanoic acid.
  • the quality of the coating was then checked by measuring the capacitance again. Only a slight increase in capacity (1-2%) compared to the previously obtained values was measured.
  • a deodorization potential of -1400 mV was applied to the gold electrode and the stability of the capacitance value was awaited. Then so much octanethiol was added that there was a concentration of 250 ⁇ mol / 1 in the cell. The following decrease in capacity after 2 hours was approximately 30% with a half-life of 45 minutes (see FIG. 4).
  • the absolute capacitance value of 7.8 ⁇ F / cm 2 was significantly larger than that of the gold electrode, which had been coated while applying the adsorption potential. After cleaning the electrode with ultrapure water and chloroform (5 seconds), it was found that the decrease in capacity was only due to physically adsorbed thiol, because the initial values of 12-14 ⁇ F / cm 2 were reached again. The physical adsorption and the subsequent cleaning step were repeated several times and always gave the same result.
  • a deodorization potential of -1400 mV was applied to the gold electrode and the setting of a stable capacitance value was awaited. Sufficient 6-mercaptohexanoic acid dissolved in the electrolyte was then added to a concentration of 50 ⁇ mol / l in the measuring cell. The reduction in capacity was about 7% after 1 hour (see FIG. 5). The applied deodorization potential has now been replaced by the adsorption potential (+300 mV). The decrease in capacity caused by the change in potential was overlaid by the immediately beginning adsorption and could not be determined. After about 2 hours the adsorption was complete (half-life 8 minutes) and the absolute capacitance (4.9 ⁇ F / cm 2 ) was comparable to a gold electrode coated in organic solution. By cleaning the electrode with ultrapure water and ethanol (10 seconds), no adsorbed thiol was removed.
  • Silicon wafer pieces with a size of 3.20 mm x 10.02 mm and a thickness of 450 microns were in a common sputtering process with two gold electrodes
  • the electrode was built up from a titanium and palladium layer (adhesion promoter, each 50 nm thick) and a covering gold layer (200 nm). The distance between the electrodes was 1.56 mm. The attachment of the contact point and the cleaning process were carried out analogously to the method described above.
  • the wafer with the two gold electrodes was attached together with an Ag / AgCl reference electrode (surface area approx. 1 cm 2 ) to a Teflon holder, which served as a lid for the measuring cell (a snap-lid glass. 40 mm x 19 mm) and an opening for the Had addition and removal of liquids.
  • the cell was filled with electrolyte (100 mM KCI; pH 6.7; approx. 3 ml) until the reactive surface of the gold electrodes and the reference electrode (Ag / AgCl) were completely immersed.
  • a magnetic stirrer ensured uniform mixing.
  • a lock-in amplifier with an integrated sine generator generated a constant sine signal with a frequency of 20 Hz and an amplitude of 10 mV.
  • Electrode El measured. The potential of +300 mV applied to electrode E2 was shifted to - 1400 mV. There were changes in capacitance at electrode El due to this
  • the measuring cell is set up and filled with fresh electrolyte.
  • the absolute capacitance values were measured again.
  • the electrode El coated with 6-mercaptohexaanoic acid had a value of 4.2 ⁇ F / cm 2 .
  • the second electrode was followed during the second coating step.
  • octanthiol concentration in the cell 150 ⁇ mol / 1
  • the absolute capacitance values of the two gold electrodes were 1.4 ⁇ F / cm 2 for electrode E2 and 4J ⁇ F / cm 2 for electrode El.

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Abstract

L'invention concerne une configuration sur la surface support de laquelle des couches moléculaires sont immobilisées de façon électriquement adressable. L'invention concerne également un procédé permettant d'immobiliser des molécules de façon électriquement adressable. L'invention concerne enfin un dispositif permettant de mettre en oeuvre ce procédé et l'utilisation de cette configuration comme capteur chimique et/ou biocapteur, notamment comme système multicapteur aux fins de déterminations chimiques, biologiques et physiques et pour des applications dans la synthèse combinatoire sur la surface limite.
EP98966157A 1997-11-21 1998-11-20 Procede de production des structures d'organisation laterale sur des surfaces support Withdrawn EP1032826A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE19751658 1997-11-21
DE19751658A DE19751658A1 (de) 1997-11-21 1997-11-21 Verfahren zur Bildung lateral organisierter Strukturen auf Trägeroberflächen
PCT/DE1998/003437 WO1999027355A1 (fr) 1997-11-21 1998-11-20 Procede de production des structures d'organisation laterale sur des surfaces support

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EP1032826A1 true EP1032826A1 (fr) 2000-09-06

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EP98966157A Withdrawn EP1032826A1 (fr) 1997-11-21 1998-11-20 Procede de production des structures d'organisation laterale sur des surfaces support

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US (1) US6458600B1 (fr)
EP (1) EP1032826A1 (fr)
DE (1) DE19751658A1 (fr)
WO (1) WO1999027355A1 (fr)

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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