MXPA00004543A - Nanoelectrode arrays - Google Patents

Abstract

An array of electrodes at the atomic or nano scale (nanoelectrodes) is built on a chip. The spatial distribution, height, width and electrochemical composition of the nanoelectrodes is varied, such that protein-specific electronic receptors are built directly on the chip with the nanoelectrodes without the use of any specific binding agents or molecules. Because of their size, a very large number of different receptors can be built as arrays on a single chip. The chip can be used to detect, characterize and quantify single molecules in solution such as individual proteins, complex protein mixtures, DNA or other molecules.

Description

NANOELECTRODE SETS This application claims the benefit of the Provisional Patent Application of the E.U.A. No. 60 / 065,373 filed November 12, 1997. TECHNOLOGICAL FIELD The present invention relates in general to methods and apparatus for detecting and characterizing simple biological molecules in solution and more specifically for detecting and characterizing individual proteins, protein mixtures, DNA and other molecules in a flake. BACKGROUND OF THE INVENTION The characterization and quantification of individual proteins or complex biological molecules is extremely important in fields as distant as medicine, forensic and military. For example, in medicine the presence and concentration of certain proteins can be used to diagnose diseases or pre-conditions of diseases. In the military, certain proteins can be used to signal the presence or absence of certain pathogens in the environment that is extremely important for example in potential germ warfare situations. The detection of individual proteins or molecules in biological samples is currently complex and generally requires sophisticated and bulky equipment.

Several technologies have recently been described to characterize certain biological molecules. In particular, success has been achieved in high density DNA chips made by Affymetrix, as originally described in PCT International Publication No. WO 90/15070. The U.S. Patent No. 5,624,537, entitled "BIOSENSOR AND INTERFACE MEMBRANE" (MEMBRANE OF INTERFACE AND BIODETECTOR), describes a receptor matrix of proteins and a single electrode. The U.S. Patent No. 5,395,587, titled "SURFACE PLASMON RESONANCE DETECTOR HAVING COLLECTOR FOR ELUTED LÍGATE" (DETECTION OF SURGICAL PLASMON RESONANCE THAT HAS COLLECTOR FOR ELIGIBLE LINKED), describes a system for measuring immobilized ligands, using a plasmon resonance detector. The U.S. Patent No. 5,607,567, entitled "PROTAMINE-RESPONSIVE POLYMERIC MEMBRANE ELECTRODE" (POLYMERIC MEMBRANE RESPONSE TO PROTAMINE ELECTRODE), describes a membrane electrode. The U.S. Patent No. 5,328,847, titled "THIN MEMBRANE SENSOR WITH BIOCHEMICAL SWITCH", describes a biodetector with a specific recognition biomolecule.

The U.S. Patent No. 4,777,019, entitled "BIOSENSOR" (BIODETECTOR), describes a biodetector for biological monomers. The U.S. Patent No. 5,532,128, entitled "MULTI-SITE DETECTION APPARATUS" (DEVICE OF DETECTION OF MULTIPLE SITES), describes test wells combined with electrodes to detect certain biological molecules. The U.S. Patent No. 4,983,510, entitled "ENZYMES IMMOBILIZED ON LATEX POLYMER PARTICLES FOR USE WITH AN AMINO ACID ELECTROSENSOR" (IMMOBILIZED ENZYMES IN LATEX POLYMER PARTICLES FOR USE WITH AN ACID AMINO ELECTRODETECTOR), describes an electrodetector with a latex polymer trap . The U.S. Patent No. 5,384,028, entitled "BIOSENSOR WITH A DATA MEMORY" (BIODETECTOR WITH A DATA MEMORY), describes a membrane biodetector with a memory module. The U.S. Patent No. 5,567,301, with the title "ANTIBODY COVALENTLY BOUND FILM IMMUNOBIO-SENSOR" (COVERAGEALLY LINKED ANTIBODY WITH FILM IMMUNOBIODETECTORA), describes an antibody biosensor. The U.S. Patent No. 5,310,469, entitled "BIOSENSOR WITH A MEMBRANE CONTAINING BIOLOGICALLY ACTIVE MATERIAL" (BIODETECTOR WITH A MEMBRANE CONTAINING BIOLOGICALLY ACTIVE MATERIAL), describes a membrane biodetector. The U.S. Patent No. 5,019,238, with the title "MEANS FOR QUANTITATIVE DETERMINATION OF ANALYTE IN LIQUIDS "(MEANS FOR QUANTITATIVE DETERMINATION OF ANALYTE IN LIQUIDS), describes a means to sequentially test the ionic concentration of fluids. The U.S. Patent No. 4,981,572, entitled "ELECTRODE UNIT AND PACKAGE FOR A BLOOD ANALYZER" (ELECTRODE UNIT AND PACKAGE FOR AN ANALYZER OF BLOOD), describes an electrode and apparatus to analyze blood. The U.S. Patent No. 4,452,682, titled "APPARATUS FOR MEASURING CLINICAL EMERGENCY CHECK ITEMS OF BLOOD "(APPARATUS TO MEASURE VERIFICATION ITEMS CLINICAL EMERGENCY OF THE BLOOD), describes an apparatus to measure multiple elements in the blood. The U.S. Patent No. 4,568,444, entitled "CHEMICAL SUBSTANCE MEASURING APPARATUS", describes an electrode for quantifying chemical substances in a solution. The U.S. Patent No. 5,281,539, entitled "IMMUNOASSAY DEVICE FOR CONTINUOUS MONITORING" (IMMUNOASSAY DEVICE FOR CONTINUOUS VERIFICATION), describes a two-stage immunoassay device. The U.S. Patent No. 5,192,507, entitled "RECEIVER-BASED BIOSENSORS" (BIODETECTORS BASED ON RECEIVER), describes a biosensor based on a polymeric film to detect opiates. The U.S. Patent No. 5,156,810, entitled "BIOSENSORS EMPLOYING ELECTRICAL, OPTICAL AND MECHANICAL SIGNALS" (BIODETECTORS USING ELECTRICAL, OPTICAL AND MECHANICAL SIGNALS), describes a thin-film biodetector. The U.S. Patent No. 5,494,831, entitled "ELECTROCHEMICAL IMMUNOSENSOR SYSTEM AND METHODS" (SYSTEM AND METHODS OF ELECTROCHEMICAL IMMUNODETECTOR), describes an immunological biodetector. The U.S. Patent No. 5,332,479, entitled "BIOSENSOR AND METHOD OF QUANTITATIVE ANALYSIS USING THE SAME" (BIODETECTOH AND METHOD FOR QUANTITATIVE ANALYSIS USING THE SAME), describes an electrode-based sensor with a biologically active receptor. The U.S. Patent No. 5,582,697, with the title "BIOSENSOR, AND METHOD AND A DEVICE FOR QUANTIFYING A SUBSTRATE IN A SAMPLE LIQUID USING THE SAME " (BIODETECTOR AND A METHOD AND DEVICE FOR QUANTIFYING AN SUBSTRATE IN A LIQUID SAMPLE USING THE SAME), describes a biosensor based on the measurement of reduction between a substrate and a dideoreductase. The U.S. Patent No. 4,908,112, entitled "SILICON SEMICONDUCTOR WAFER FOR ANALYZING MICRONIC BIOLOGICAL SAMPLES" (OBLEA SEMI - SILICON CONDUCTOR FOR ANALYZING BIOLOGICAL MICRONIC SAMPLES), describes a microspapillary separation device with detector capabilities. The U.S. Patent No. 5,409,583, titled "METHOD FOR MEASURING CONCENTRATIONS OF SUBSTRATES IN A SAMPLE LIQUID BY USING A BIOSENSOR" (METHOD FOR MEASURING SUBSTRATE CONCENTRATIONS IN A SAMPLE LIQUID WHEN USING A BIODETECTOR), describes a two-stage biosensor. The statutory invention of the U.S.A. H201, with the title "BIOSENSOR FROM MEMBRANE PROTEINS RECONSTITUTED IN POLYMERIZED LIPID BILAYERS", describes a method to incorporate and use cell membrane proteins in biosensors. The technologies described above are generally used for the detection of a single type or a few different types of molecules. None of these technologies is particularly adapted to allow a very large number of different types of proteins, protein variants or other biological molecules to be detected and quantified simultaneously in a single flake. Furthermore, nothing in the prior art provides a convenient technology for directly constructing protein-specific electronic receptors in a flake, without the use of any biological building agents, synthetic probes or complex micro-structures such as test wells. Here we describe a novel, smaller, faster and more cost effective technique to detect, characterize and quantify individual proteins or other complex molecules in a flake. The technology described here can also serve as a new method for DNA sequencing. COMPENDIUM OF THE INVENTION In one aspect, the present invention provides a detector that is capable of distinguishing between different molecular structures in a mixture. The device includes a substrate in which nanoscale binding sites are manufactured in the form of swarms of multiple electrodes. Each binding site includes nanometer scale points that extend over the surface of a substrate. These points of preference are nanoelectrodes that are spatially configured to provide a three-dimensional electrochemical junction profile that mimics a chemical binding site. In this way, the binding sites have selective affinity for a complementary binding site in a target molecule or for the target molecule itself. In one aspect, the binding sites are arranged in a set on the substrate. In one aspect, the spatial and electrochemical profiles of each site in the set are identical and provide an assay for a simple target molecule. In another aspect, regions of the nanoelectrode array carry clustered sets of electronically and / or spatially distinct binding sites for simultaneous detection and quantification of several molecular species. In yet another aspect, the materials used for the electrodes and surrounding surfaces are chosen based on preferred intrinsic electrical and chemical properties. The nanoelectrode assembly can be included in a chamber that can retain fluid. Several assemblies can be used in a single chamber and several different chambers can be used in a single flake. In yet another aspect, the nanoelectrode array and the chamber are connected at least to a microfluidic separation and delivery system such as a micro-capillary, which allows both the delivery and separation in size and electrical properties of proteins or other molecules to analyze. In another aspect, a microcontroller or microprocessor is provided to analyze signals from the nanoelectrodes and / or to synchronize and control the fluidic separation of the molecules or proteins. In another aspect, the flake with the nanoelectrode assemblies is associated with a system for electronic temperature control, such as a thermoelectric device having a thermistor to vary the kinetics of binding or the electrochemical affinity of the molecules with certain nanoelectrodes, as well as the kinetics of flow and separation of molecules. In another aspect, the nanoelectrodes are interspersed in a linear microtube to sequence DNA. Thus, an object of the present invention is to provide a novel and rapid method for analyzing small biological molecules in solution such as proteins and for sequencing DNA by using semiconductor chip technology with extremely high packing densities. A further objective of the present invention is to ensure that all the flake can be easily integrated into devices for automated analysis of proteins, DNA or other molecules. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a diagrammatic perspective view of a set of nanoelectrodes showing different swarms of nanoelectrodes. Figure 2 is a diagrammatic view in lateral elevation of a specific protein receptor and its corresponding protein. Figure 2A is a lateral elevation cross section of a specific protein receptor and its corresponding protein. Figure 3 is a cross-sectional, side elevation view of a nanoelectrode assembly within a microfluidic tube showing entrapment of a specific protein in its corresponding nanoelectrode receptor. Figure 4 is a cross sectional view, in diagrammatic side elevation of a microtube with a linear nanoelectrode array for detecting DNA. Figure 5 is a cross section of an integrated flake with nanoelectrode assemblies, a microfluidic delivery system and associated electronic components. Figure 6 is a lateral elevation cross-section of a nanoelectrode receiver, showing the electric field that is interrupted or modified by joining a specific molecule to the receptor.

Figure 7 is a view of a cantilevered nanoplate with several swarms of identical nanoelectrodes. DETAILED DESCRIPTION OF THE PREFERRED MODALITIES The present invention is based in part on the fact that recent advances in technology such as the use of scanning tunneling microscopy (STM) Scanning Tunneling Microscopy), have shown that ultra-small structures of a simple atomic layer or of a few, can be built on a semi-conductor surface such as silicon. Due to the size of these structures, they are generally referred to as nanostructures (one nm nm = 10"9m, 1 Angstrom or Á = 10" 10m). These structures can be as small as a few Angstroms in diameter that are well below the Stokes radius of a small protein (which is approximately 25-35Á). Since these structures can be constructed using different chemical elements (or the voltage applied to the structure can be varied selectively) and the spatial distribution, width, height and shape of the structures can also be varied, these structures can be constructed in swarms to serve specifically as " Molecular electrodes "whose electrochemical properties and spatial distribution can be made to correspond precisely with the external three-dimensional shape and electrochemical properties of molecules, preferably biochemical and more preferably proteins. Therefore, each of these swarms can serve as "receivers" (or detectors) of individual electronic proteins. Since a very large number of these molecular electrodes can be placed in a single flake, the resulting sets, here called "nanoelectrode arrays" can be used to detect, characterize and quantify very different proteins in a single flake. In a variation of the technology, the flake can also be used to sequence DNA. Now with reference to Figure 1 of the drawings, the microelectronic molecular detector 20 is seen to have a substrate 22 in which a set of binding sites or swarms 24 is formed. The substrate 22 can comprise any number of materials such as silicon , germanium, gallium arsenide or other semiconductors. Now with reference to Figure 2 of the drawings, a bonding site 24 is illustrated in more detail having multiple electrodes 26a, 26b and 26c that are spatially distributed to form a pattern. In this way, it can be seen that each electrode 26a, 26b and 26c in this particular embodiment is laterally spaced from the adjacent electrode and rises at different heights apart from the main surface 28 of the substrate 22. It will be appreciated that through molecular modeling and data empirical, the topology of the binding sites and the electrical charge are sized to the extent, to provide the topographic and electrical properties required to selectively recognize and bind a complementary region of a target molecule. As best illustrated in Figure 2, protein 30, which has a defined form specific to that protein, is connected to a nanoelectrode swarm determined from three nanoelectrodes 26a, 26b and 26c. As will be explained more fully, each nanoelectrode may have slightly different electrochemical properties due to different charges and / or chemical compositions. These individual electrochemical properties correspond not only to the electrochemical affinities of the amino acids or atoms present in the slots of the protein, but also complement the shape of the slit itself. In this way, when a molecule tending the appropriate complementary profile is linked to the "receiver" 24 bridging the space between the electrodes, a change in electrical potential occurs which can be verified through appropriate circuits to provide an indication of the presence of the target molecule. In the most preferred embodiments of the present invention, the binding sites 24 have nanoscale geometries. As illustrated in Figure 2, the distance from the main surface 28 to the top of the electrode 26b is 1.9 nanometers, the width of the electrode 26b is 0.7 nanometer and the distance between the electrodes 26b and 26c is 1 nanometer. In general, each electrode will typically be between 0.2 and about 3 nanometers in height and from about 0.2 to about 2 nanometers wide. As it is used here, "nanoelectrode" will include atomic-scale structures as well as nanoscale, that is, from 2Á to 5 nanometers. Also typically from about 2 to about 8 separate electrodes are given in each swarm 24. The electrodes 26a, 26b and 26c can be formed from a number of materials, either intrinsic or adulterated such as gold and platinum and copper and other electrometals. Gold is particularly preferred. It may also be convenient to form the electrodes of a material and coat the outer portion with a different material, for example gold coated with zinc oxide or gold coated with a thiol group. Each of the electrodes can be connected separately to a power source by small regions or conductive wires that can be formed of gold. In Figure 2A, the individual conductive layers 34a, 34b and 34c, are illustrated to electrically connect their respective electrodes 26a, 26b and 26c. The dielectric layers 36 electrically insulate the individual conductive layers and the dielectric sheaths 38 electrically insulate the individual electrodes. It will be appreciated that different potentials can be applied to the various individual electrodes and that electrodes of different swarms can be electrically linked to a single layer, for example layer 34a. It will be appreciated that the various layers can be formed using conventional thin film manufacturing techniques such as CVD, thermal growth and ion implant. It has recently been shown that electric "wires" of simple atoms can be constructed (see for example the article by Leo Kouwenhoven "Single-Molecule Transistors", Science, Vol. 275, pages 1896-1897, March 28 , 1997, the entire description of which is incorporated herein by reference). The wires can be deposited in a number of different ways as part of the manufacturing process of micro-flakes, before deposition of the nanoelectrodes. The nanoelectrodes can be deposited directly into the flake by Scanning Tunnel Effect Microscope (as described by Kolb et al., Science, Vol. 275, pages 1097-1099, February 21, 1997, the entire description of which is incorporated herein by reference. ). A number of other methods of manufacturing flakes are possible such as different lithography techniques, etc. In another aspect, the nanoelectrodes are not connected to any electrical wire or conductive layers. In this case, the binding of the protein or other molecule simply depends on the shape and chemical properties of the individual nanoelectrode swarms. Detection of the connection of a given molecule with a determined swarm can then be achieved by means other than electrical, for example by a positional fluorescence reader x-y, highly accurate, similar to that used for the technology of DNA lacquer or by resonance. In the case where the nanoelectrodes are not connected to wires (ie they are not "live" electrodes), in some applications the nanoelectrodes can interconnect in a determined swarm. In this case the swarms will comprise interconnected peaks and slots and these will form a larger structure (ie from 1 to >10 nanometers). This structure can be adjusted to the extent, either to correspond precisely with the current biological receptor of the target molecule or allow the whole molecule to fit in a "three-dimensional" receiver, which would correspond to at least a third of the form 3- Total D of the molecule. In some cases and depending on the total shape of the molecule, the receptor that is constructed may not necessarily include a site that corresponds to the current biological receptor of the target molecule. Several types of binding or adsorption of the molecule to the nanoelectrode receptor are possible, depending on the chemical composition of the nanoelectrodes, the voltage and the chemical to be measured. Binding forces can include covalent bonding, electrostatic bonding, hydrogen bonds and van der Waals bonds. Depending on the type of detection required, the individual nanoelectrodes of individual swarms do not necessarily need to be composed of different electrometals, since both the spatial distribution and the height of the nanoelectrodes can be varied and these two variables can be sufficient for detection of specific molecules in certain applications. In some applications, each nanoelectrode can be selectively charged in a given swarm, allowing the electrophysical property of the nanoelectrode to be varied. The entire detector can be constructed using a computer controlled operation, wherein the spacing, height, width and composition of the nanoelectrodes can be made to exactly correspond to the three-dimensional shape and respective electrochemical properties of a select molecule. In addition, since the position of the nanoelectrode swarms corresponding to a specific receptor for a specific molecule is determined during the manufacturing process, this position information can be used to detect connection or binding. For example, a large set of nanoelectrodes can be constructed with very different swarms, ligated in solution can be allowed, then the set can be read using a highly accurate x-y reader in a manner similar to DNA lacquer. Computer-controlled manufacturing of the nanoelectrodes also allows identical copies of the flake to be made. It will also be appreciated that the geometries that are constructed on the surface of the flake can be made to correspond exactly to the corresponding image of a crystallized protein surface that is taken from x-ray diffraction studies. Therefore, swarms of nanoelectrode arrays can be constructed directly using crystallographic data and the resulting surfaces in the flake will favor the specific crystallization of protein in certain sets. In another aspect, since multiple identical receptors can be built on the same flake, this technology can be used not only to detect specific molecules, but also to estimate precisely the amount of these molecules present in the sample, when measuring the speeds of union in swarms identical With reference to Figure 3, two sets of partial nanoelectrodes facing each other and forming the microchannel or nanotube 60 are illustrated, allowing the flow of small molecules such as through protein 70. If the protein 70 corresponds to the shape of a receptor composed of electrodes 74, 78 and 82, the physical binding of the protein will cause a minute minute change in the electrical signal, which can be measured simultaneously in all the nanoelectrodes. The intensity of the electrical signal can be modified, for example, by adding a conductor to the carrier solution for the molecules that need to be studied. Alternatively, the nanoelectrodes themselves can be charged with a small current, which would change before connection of the given molecule. Depending on the electrochemical properties of the nanoelectrodes and the analyte, the temperature and the expense of flow, the union can last only a fraction of a second or last longer. The retention time itself is another important variable that can be used to detect and quantify the types of molecules present in the sample. In some applications, the micro-channel 60 can form a part of a network of channels of different and specific sizes, corresponding to the sizes of the proteins to be measured. Each of these channels can be equipped with molecular sieves, allowing only certain proteins or molecules of a size to pass through. The channels themselves can also serve as a means for separating molecules and supplying them to determined detector chambers with sets of nanoelectrodes that are specifically made to measure certain classes of proteins or molecules of determined molecular weights. In this case, each of the sets will have nanoelectrodes with sizes corresponding to the sizes of the proteins to be measured. As part of this network of channels, specific cameras can be added with specific functions such as a camera to lyse cells. Other chambers can be filled with specific reagents that can be used as required. In other applications, each of the micro-channels is equipped with only one or a few swarms of nanoelectrodes and the protein mixture is circulated through each of the channels. With the help of a microcontroller or a microprocessor that controls the flow rate in each microchannel, the signals from each of the nanoelectrode swarms are then measured by combining the power of the following variables for detection: protein separation rates (based on in the size and charge of the proteins) and retention time in each determined swarm (based on the shape and electrochemical properties of the molecule). Undoubtedly, the more a given molecule corresponds to a given receptor, the longer it will bind. It will be evident that the sophisticated control and measurement of the electrical signals in each nanoelectrode (as well as the control of all other variables such as sample flow rate, temperatures, etc.), can only be done with the help of a microcontroller or a microprocessor. Now with reference to Figure 4, a nanoassembly of electrodes 90 is constructed in a linear microtube 100, with the spacing and electrochemical composition of the nanoelectrodes that are varied such that they correspond exactly to the distance between determined base pairs of a linear portion. of DNA or RNA 110. In this case, the nanoelectrodes are constructed using only two variables: precise spacing and electrochemical composition (not height) that favor specific binding of position of specific base pairs of DNA or RNA to corresponding nanoelectrodes. The principle that is applied here is that DNA is known to behave like a linear molecule, when it is circulated in a microtube and that this flow rate can be controlled and measured accurately. In addition, the distance between 10 base pairs of DNA that is precisely 34A, the nanoelectrodes can be spaced precisely in multiples of 3.4A as illustrated in 120. By varying the spacing and loading and / or composition of the nanoelectrodes and by measuring changes in Conductance over time in nanoelectrodes placed sequentially, a whole sequence is created based on the synchronization of position-specific nanoelectrode signals. The entire DNA (or RNA) sequence is then reconstructed with the help of a microcontroller (or microprocessor) that can also control the flow rate in the microtube. ANALYSIS OF PROTEIN VARIANTS Mutations or other changes in DNA result in substitutions of amino acids in the protein. These substitutions in turn result in conformational changes in the protein and can result in proteins that are already non-functional or have different properties. Since the three-dimensional (3-D) structure of proteins can now be accurately inferred based on x-ray crystallography or nuclear magnetic resonance (NMR), the 3-D forms of the protein variants can also be generated using the same method. Therefore, the complete spectrum of protein variants for certain classes of proteins can be measured and quantified using the nanotechnology described above. This is because the conformational changes of each protein variant can be represented by a swarm of determined nanoelectrodes that vary in the shape, distribution and electrochemical properties of the nanoelectrodes. In fact, the construction of the sets can be controlled by computer and link the information corresponding to the putative 3-D structure of proteins of interest (and its variants) with the micro-fabrication of all the corresponding receptors in the flake. By measuring and quantifying these variants as described above, this approach represents a powerful alternative for detecting DNA sequencing, since all possible mutation products of certain genes that are expressed can be measured directly on a flake. Another advantage is that the flake will be completely reusable. In addition, given the extremely high density of the nanoelectrode arrays that can be constructed in a single flake, all spectra of protein variants for many genes can be measured immediately in the same flake. In fact, with a refinement in technology, all existing human proteins and their variants can theoretically be measured in a single 1 cm2 flake and the number of receivers that can be built in this flake will theoretically exceed one billion, which is an improvement of one thousand times against any existing technology.

SEPARATION OF PROTEINS As indicated above, the separation of molecules can be achieved by circulating the molecules in extremely small tubes (micro-capillaries, micro-channels or nanotubes), where the smaller molecules travel faster than the larger ones that are retained by friction and weak bonding interactions with the surfaces of the tubes. The result that is achieved is equivalent to electrophoresis but with the advantages of speed, cost and reuse of microthreads. Now with reference to Figure 5, the micro-channel 130 is illustrated with a sample feed gate 132 and a long loop circulating within an optional reagent micro-chamber 134, the same connected with an optional feed gate 136 The micro-channel 130 separates biological molecules by size and charge, while the micro-chamber 134 allows the selective feeding of a reagent or external solution. The flow and on / off position of each micro-channel junction can be controlled electronically either by an external micro-pump (not shown), by thermocapillary action or by an electric potential change. After entering the micro-chamber 134, the analyte then successfully circulates to the micro-chambers 138a, 138b, 138c and then 138d, each containing different sets of nanoelectrodes with swarms of nanoelectrodes of varying sizes and densities. In this particular design, the nanoelectrode assemblies are fabricated immediately adjacent to microelectrically multiplying components or control area 140 itself connected to an interface 142. After reacting with successive nanoelectrode arrays in successive microcameras, the sample exits through the gate 146. The micro-channels and micro-chambers can either be etched on the silicon surface itself or can be manufactured separately on a surface of a material such as glass, diamond, plastic or similar, which are then connected to the silicon surface. This design can be varied in very different ways and Figure 5 illustrates just one of the many possible combinations of micro-channels, nanoelectrode arrays and microelectronic assemblies that can be fabricated in a chip. As indicated above, a chamber that allows the lysis of the cells or virus to be analyzed can also be included in the flake. Also, it should be noted that the directional flux in the micro-channels can be reversed and that each connecting micro-channel can be selectively opened or electrically closed. Therefore, when the test is complete the entire system can be heated to allow denaturation of the protein (and / or the potential in the nanoelectrodes can be reversed), then the system can be flushed with a solution to clean the nanoelectrode assemblies and allow reuse of the lasca. Therefore a complete and integrated protein detection and separation system can be built in a single flake. An important aspect of combining sets of nanoelectrodes, micro-channels and a microcontroller (or a microprocessor) is that the separation time (from the injection of the sample into gate 132 at the time of first detection) and the length of retention in certain Nanoelectrode receptors are important variables for characterizing proteins or individual protein variants. For example, the system can be calibrated by injecting known proteins, then known mixtures of proteins, before injecting the sample to be tested. The time it takes to reach a given nanoelectrode receiver and the length of binding in different electronic receptors would be specific for proteins or specific protein variants and the specific signal profiles for each protein can then be stored in memory and compared with those of the sample to try. While Figure 5 illustrates an integrated design, it is evident that the protein separation component and electronic components can also be externally collocated and that the flake can be as simple as having a simple nanoelectrode array, circumscribed in a single chamber with a interphase This flake (which can be disposable) can then be inserted into a larger module with the above components. Also, as indicated below, other detection methods can be used and the design of the flake will be changed accordingly. DETECTION There are many ways in which the binding or adsorption of the analyte in the set of nanoelectrodes can be detected. Now with reference to Figure 6, one way to detect the signal due to adsorption in the nanoelectrode array is by electrical signal. In this case, at least one of the electrodes in each swarm of a given assembly is used as a "source" 160, while the rest of the swarm 165 is used as a "collector". When an analyte, say a protein, is adsorbed, it changes the current flow (peak amperes) as illustrated in Figure 6. The electrodes are isolated by an oxide layer 170. Unwanted effects of electric current can be avoided when using an AC approach. Now with reference to Figure 7, the second approach to detecting the joint is by using a resonance approach. In this method, a nano-structure is constructed. For example, the nanoplate 180 with a smaller dimension than a miera, is constructed. This structure can be autonomous or it can be cantilevered. Identical sets of nanoelectrode receivers 24 are then manufactured on this surface. The structure is designed to have resonance frequency in the low MHz to GHz region. As the analyte circulates these structures, it spends a longer time in the cantilevered structure if they have a structure that is complementary to the structure of nanoelectrodes. In other words, the analyte molecules are subjected to collision with the nanoplate. If there is any complementary nature between the analyte and the substrate, the analyte will spend more time on the surface during collision. This can be detected optically by having a laser diode radiate in the structure and detect the reflected signal using a position sensitive photodiode. The AC signal in the photodiode shows the resonance response of the structure. The larger the signal, the higher the concentration of bound biological molecules, that is, the higher the concentration of the molecule in the solution. Other detection techniques such as capacitive, piezo resistive, piezoelectric, with electron tunnel effect, etc., can also be used. The structure can be excited in resonance response by mechanical means using a piezoelectric element. In this technique, a nanoplate structure is connected to a piezoelectric material that can be vibrated using an AC signal. In resonance, the structures oscillate with maximum amplitude. I could also get excited in resonance by modulating in laser diode using square wave energy pulses. Since square waves contain all Fourier components, there will be a component that corresponds to the resonance frequency of the structure. Since these nanoelectrodes can be constructed in geometric structures with an extremely small thermal mass (for example nanoplates have a thermal mass in the order of many pico grams or less), they can be heated and cooled in the time frame from micro to milliseconds. This fact can be used to adsorb and desorb analytes in a periodic manner. However, when there is a complementary structure between the surface and the analytes, the desorption time scale will be different. USE OF AN EXTERNAL DETECTOR In another detection application, all the flake that can be allowed to react with the sample is placed in a x-laser reader and in the form similar to DNA flake. In this case, the flake is incorporated in a highly accurate support to ensure accurate position reading of each swarm. Detection by fluorescence can be effected, for example after reaction of the samples linked to the swarms with a fluorescent molecule or with labeled antibodies. Detection can also be performed by other means such as laser ionization-desorption mass spectrometry. CONSTRUCTION OF NANOELECTRODES The nanoelectrode assemblies can be built on a semi-conductor substrate that is adulterated by nanolithography, using scanning or scanning probes. In this approach, metal swarms are already deposited from a solution or by field evaporation from an STM / AFM tip. Since the electric field between the tip and the substrate is very high (109 V / m), many metals can be evaporated in the field. In solution, many metals can be deposited electrochemically on a substrate. The surface of the semiconductor can be oxidized to be an insulator. Nanometer-scale lines and trenches can be fabricated on a semiconducting surface using an STM tip in a rust-etched solution, producing a trench. The depth of the trench depends on the time spent by the tip at that site and the voltage at the tip. Therefore, not only nanoelectrodes can be constructed by deposition but can also be constructed by etching. The ditches can also be used to make the channels to separate the proteins as previously instructed. It will also be noted that nanotransistors can be built directly in the flake, to facilitate detection and increase the density of the detectors. The nanotransistors can be constructed before the deposition of the nanoelectrodes as a sub-layer in the total chip manufacturing process or placed in an adjacent part of the chip. The principles described above illustrate the wide variety of applications that are possible in the micro-fabrication and applications of nanoelectrode assemblies. For example, the entire system, from sample feeding to detection with output signals sent to an external device such as a monitor, can be built in a single flake, using micro-channels (for separation and sample supply), miniature ion pumps, detection sample, an interconstructed microcontroller, a method for temperature control, etc. This flake can be inserted into a measuring device, for example for use in a medical office or in field detection. If a very large set of nanonsensors is used, it may be preferable to use a microprocessor or several microcontrollers to control the functions described above. In some applications large assemblies can be used with an external laser reader. In this case, the assembly can be used in a similar way to the DNA flake, where all the flakes are allowed to react with the entire sample, washed and then reinserted into an external reader. Using this approach the flake can be constructed in a convenient handling cassette. While the invention has been described with respect to a specific embodiment for a clear and complete description, the appended claims shall not be limited in this way but shall be constructed incorporating all modifications and alternate constructions that may occur to them. a person with skill in the specialty that falls substantially within the basic teachings established here.

Claims (37)

1. CLAIMS 1. A detector for biological molecules, the detector is characterized in that it comprises: a substrate; an electrode that has the ability to bind a preselected biological molecule, the electrode is between approximately 10"9 and 10" 10 meters in height and width.
2. 2. The detector according to claim 1, characterized in that the electrode is a plurality of electrodes.
3. 3. The detector according to claim 2, characterized in that each of the electrodes has an identical chemical composition. .
4. The detector according to claim 2, characterized in that at least one of the electrodes has a chemical composition that is different from the other of the electrodes.
5. 5. The detector according to claim 1, characterized in that the electrode has an outer coating.
6. 6. The detector according to claim 2, characterized in that each of the electrodes has a chemical coating.
7. The detector according to claim 6, characterized in that each of the coatings has the same chemical composition.
8. 8. The detector according to claim 6, characterized in that at least one of the coatings is different from the other of the coatings.
9. The detector according to claim 2, characterized in that the height of at least one of the electrodes is different from the height of the other of the electrodes.
10. The detector according to claim 2, characterized in that the width of at least one of the electrodes is different from the widths of the other electrodes.
11. The detector according to claim 2, characterized in that the electrodes are laterally spaced apart from each other on the substrate.
12. 12. The detector according to claim 2, characterized in that the electrodes are arranged in swarms in the substrate.
13. The detector according to claim 2, characterized in that the electro-chemical properties, width and spacing of the electrodes complement each other and link a site in the biological molecules.
14. 14. The detector according to claim 1, characterized in that the electrode is connected at least to an electrically conductive nanowire.
15. 15. The detector according to claim 2, characterized in that the electrodes are connected to nanowires.
16. 16. The detector according to claim 1, characterized in that it also comprises an interface that connects the detector to a control system.
17. 17. The detector according to claim 2, characterized in that the swarms are spaced apart to form an assembly.
18. 18. The detector according to claim 1, characterized in that the biological molecules are proteins.
19. 19. A detector for proteins, the detector is characterized in that it comprises: a micro-capillary tube; a plurality of electrodes arranged in the tube, the electrodes have the ability to bind a preselected protein, the electrodes are between approximately 10"9 and 10" 10 meter in height and width.
20. 20. The detector according to claim 19, characterized in that it also comprises a microcontroller.
21. 21. The detector according to claim 20, characterized in that it further comprises a system for regulating the temperature of the detector.
22. 22. A detector for biological molecules, the detector is characterized in that it comprises: a substrate; a set of cantilevered micro structure in the substrate; at least one electrode arranged in at least one of the cantilevered micro structures.
23. 23. The detector according to claim 22, characterized in that it also comprises a laser for determining the concentration of biological molecules linked to the electrode.
24. 24. The detector according to claim 23, characterized in that it also comprises a piezoelectric detector for the concentration of biological molecules linked to the electrode.
25. 25. A method for sequencing nucleic acids, characterized in that it comprises the steps of: providing a detector, the detector having a substrate in which the plurality of electrodes are arranged, each of the electrodes being between approximately 10"9 and 10" meter height and width; contact the electrodes with a solution containing nucleic acids; The electrodes have the ability to bind at least some of the nucleic acids.
26. 26. The method according to claim 25, characterized in that the nucleic acids are DNA and wherein the electrodes are spaced apart to complement and ligate to base pairs of DNA of a linear DNA molecule.
27. 27. The method according to claim 25, characterized in that the detector includes a microtube in which the electrodes are arranged.
28. 28. The method according to claim 25, characterized in that it also comprises a flow control system and laser detector.
29. 29. The method according to claim 25, characterized in that it further comprises a microcontroller and an exhibitor.
30. 30. The method according to claim 25, characterized in that the nucleic acids are RNA.
31. 31. The method according to claim 2, characterized in that it also comprises a support structure for the substrate, the support structure is adapted to be received in a fluorescent laser reader x-y.
32. 32. A silicon chip for detecting individual proteins, comprising at least one detector manufactured with precision at the Angstrom level, wherein the surface of the detector exactly complements the three-dimensional shape of a given protein.
33. 33. The invention according to claim 32, characterized in that the detector is made of a single metal.
34. 34. The invention according to claim 32, characterized in that the detector is made of different metals.
35. 35. The invention according to claim 32, characterized in that the detector forms a specific protein receptor.
36. 36. The invention according to claim 32, characterized in that the detector is elaborated from information derived from x-ray diffraction studies.
37. 37. The invention according to claim 32, characterized in that the detector is made from information derived from nuclear magnetic resonance studies.