TW201124722A - Preparation method of antibody probe chip with electron-conducting molecule - Google Patents

Abstract

A method is disclosed for preparation of an antibody probe chip having organic electron-conducting anchoring molecules covalently bound to the metal film on the chip substrate. The antibody probe with covalent bonding organic electron-conducting anchoring molecules allows for fast and efficient molecular sample detection, characterization and measurement via binding to corresponding target molecule in a sample. Covalent bonding of the probes with the organic electron-conducting anchoring molecules on the chip substrate improves the stability and conductivity of the antibody probe chip. Due to such stability improved, a test chip can be washed with adequate solution without substantial lost of molecules already bound to the probes. Due to such electronic conductivity improved, the probes bound to the organic electron-conducting anchoring molecules can be applied in field of electrosensing chips. Thorough cleaning of the chip reduces to minimum the interference from debris to the test, which leads to improved precision in the detection and characterization results. Moreover, an electron-conducting antibody probe chip comprises a substrate having coated with a surface thereof a layer of metal film. First end of each of a number of organic electron-conducting anchoring molecules being directly bound to the metal film on the substrate, or being indirectly bound to the metal film with a linking compound, and an electron-conducting antibody molecule as the probe, which has been modified with organic electron-conducting molecules, is covalently bound to the second end of a corresponding one of the elongated anchoring molecules.

Description

201124722 VI. Description of the Invention: [Technical Field] The present invention relates to an electron-conducting antibody probe chip, and more particularly to a method for fabricating a basic antibody probe wafer on a metal substrate thereof. Covalent bonding (organic electron-conducting anchoring molecule), which is also covalently bonded to the organic-conducting molecule (electron-conducting • molecule) Therefore, the stability of the wafer substrate surface-immobilized probe and the ability of electron conduction can be increased, and the method for fabricating the antibody probe wafer for rapid measurement of the sample inductance can be quickly and sensitively performed. [Prior Art] An antibody probe chip (antibody probe chips) can be prepared by immobilizing a selected antibody probe with an anchoring molecule on a substrate having an extended surface. ). The antibody probe wafers designed for a wide range of applications are used for detection sensing purposes of specific targets. For example, in the field of basic medicine and clinical medicine, an antibody probe wafer, which is generally known as a biochip or a biological sensor chip, can quickly perform a biological sample. Detect and characterization. In medical use, the target for detection and judgment may be a biomolecule present in the sample, virus, 201124722 bacteria or cells. Among other industrial uses in the non-medical field, it may involve qualitative qualitative quantification of industrial tests such as food or environmental sampling.

In the use of biochips or biosensor chips, an antibody probe immobilized on the surface of the substrate can be affinity-activated with a target in the test sample solution. Use (affinity reaction). Specific biomolecules can be detected based on the results of the reaction. The probe immobilized on the surface of the substrate may be an antibody or a fragment thereof, such as immunoglobulin G, immunoglobulin M, immunoglobulin A, and immunoglobulin A (Immunoglobulin A). Immunoglobulin E (Immunoglobulin D) or its immuno-binding fragments, or other affinity reaction or catalysis with the target to be tested a biomolecule of a virus, cell or tissue surface. Such a substrate is coupled with an immobilized antibody or a fragment thereof as a probe (probe> for biosynthesis in a medical sample of a biomedical sample (ie, a target to be analyzed (targe cut occurs for affinity, borrowing By examining the binding concentration or binding constant of the interaction between the two, a basic data diagnosis and experimental parameters can be provided. High affinity between the two^ High binding capacity and biostability are very important features in the detection of biochips. There are a variety of bioassays that require biomolecules such as proteins to be directly immobilized on the substrate. For example, Western blotting adsorbs proteins on polyvinylidene fluoride (PVDF) or nitrocellulose membrane substrates as probes, which can interact with surface functional groups on antibodies or antigenic proteins. Function to measure whether the sample contains such protein. In addition, another example is enzyme-linked immunosorbent assay The method (enZyme-linked immuno-sorbent assay, ELISA) is to adsorb a specific antibody (or antigen) on a polystyrene substrate, and then expose the substrate to the sample. If the sample contains These antibodies (or antigens) have high affinity specific antigens (or antibodies) that are combined with them to be detected by light output. For example, Rampal et al. S.  Pat.  No.  6,013,789,"Covalent attachment of biomolecules to derivatized polypropylene supports''^ The polypropylene film is first aminated, and then reacted with the polynucleotide containing the phosphoricidazolide at the end, thus, the polynucleotide Covalent attachment to polypropylene can be achieved via phosphoamidate bonding. ^ In addition, the use of self-assembly of molecules on the surface of indium tin oxide (ITO) to form self-assembled monolayers can also provide a new interface for exploring certain biomolecule specificities. Redox reaction. The organic ruthenium may be reacted with a substrate having a hydroxyl group on the surface (for example, ruthenium dioxide) to form a single or double layer organic ruthenium film. For example, Chrisey et al. S.  Pat.  No. In the case of "Selective attachment of biomolecules to patterned self-assembled surfaces", No. 5,688,642, an organic sand compound 201124722 having two active groups is proposed, which uses a reactive group at one end to form a linkage with a hydroxyl group on the surface of the substrate, and then the activity at the other end. The base links the biomolecule so that the biomolecule can be immobilized on the substrate. Although these methods have been widely used, there are disadvantages such as cumbersome and time-consuming non-conductivity of biomolecular probe preparation and unstable optical output, and limitations on commercial mass production. These biomolecular probes are often prepared by using a long chain of alkyl long-chain compounds to link substrates and probes, which are difficult to perform π-conjugated system electron transfer and conduction properties, and are difficult to apply in electricity and electronics. Biosensing technology. Moreover, due to the immobilization of biomolecule probe molecules on commonly used substrates such as PVDF, nitrocellulose or microscopic glass, the physical adsorption effect that is usually relied on is an instability. And a time consuming process. The entire immobilization process typically requires at least six to eight hours of incubation time. A typical treatment method is to first add a biomolecule probe to a wafer substrate and place the wafer. After a physical adsorption reaction between the biomolecule probe and the substrate for a certain period of time (about three to four hours), the ® procedure is repeated several times. After the cleaning, the blocking reagent is added, and after standing for a long time (usually about three to four hours), the wafer can be used as a biomolecular probe wafer. SUMMARY OF THE INVENTION Therefore, there is a need to provide a biomolecular probe chip, particularly an antibody probe chip, having a covalently bonded electron-conducting anchoring molecule. 201124722 molecules), or antibodies coated with organic-conducting molecules, which have the ability to conduct electrons with conduction electrons, which are simple and fast to prepare, and have high stability in storage and use. And specificity, and can quickly and accurately produce the electrical test of the sample. In order to achieve the foregoing and other objects, the present invention provides a method for fabricating a basic antibody probe wafer, comprising: a covalently bonded organic electrically conductive anchor molecule comprising: a substrate having a metal film layer disposed on a surface thereof; A plurality of organic conductance anchors, molecules having a cyclic chain molecular structure with an electron transfer capability of a π-conjugated system, mainly oligophenyl, oligothiophene, oligopyrrol, and oligosaccharide Oligopyridine cyclic chain molecular structure ^ each of these organic conducting anchor molecules, each of which is directly covalently bonded to the metal thin film layer at one end of the cyclic chain molecular structure, or through a coupling The molecule is indirectly covalently bonded to the metal thin film layer and a plurality of probe molecules, each of the basic antibody probes is covalently bonded to the anchor molecules, and the corresponding anchor molecules are opposed to the cyclic chain molecules. a second end of the first end of the structure. The invention also provides a method for fabricating a conductive antibody probe wafer, wherein a plurality of organic conducting molecules, substantially having an extended and substantially circular chain molecule, are covalently bonded to the basic antibody probe, each of which Each of the electrically conductive antibody probes is covalently bonded to the second end of the first end of the organic chain of the molecular structure. The invention also provides a probeless semi-finished wafer substrate manufacturing method for an antibody probe wafer, which can be pre-stored to couple the antibody probe wafer to detect the probe molecules required for the detection of 201124722, the probeless semi-finished wafer The substrate comprises: a substrate, a surface of which is covered with a metal thin film layer; and a plurality of organic conductive anchor molecules, the cyclic chain molecular structure having an electron transfer capability of the π conjugate system, each of which is determined Each of the anchor molecules is directly covalently bonded to the metal thin film layer by one end of one of its cyclic chain molecules, or indirectly covalently bonded to the metal thin film layer through a linking molecule; and it is opposed to the ring The second end of the first end of the chain molecular configuration can be covalently bonded to the probe molecule if necessary to form the molecular probe crystal. The invention also provides a method for fabricating a conductive antibody probe wafer, wherein a plurality of organic conducting molecules, substantially having an extended and substantially circular chain molecule, are covalently bonded to the basic antibody probe, each of which The electrically conductive antibody probes are each covalently linked to the second end of the first end of the cyclic chain molecular structure corresponding to a certain anchor molecule in the organic conductance anchor molecules. The invention also provides a method for fabricating an antibody probe wafer, the method comprising the steps of: coating a metal film layer on a surface of a substrate; and further comprising a plurality of organic molecules having substantially extended and substantially cyclic chain molecules The electrically conductive anchor molecule, wherein the first end of each of the cyclic chain structures is directly covalently bonded to the metal thin film layer, or indirectly covalently bonded to the metal thin film layer through a linking molecule. The method of the present invention may further covalently bond an antibody probe to each of the anchor molecules corresponding to the second end of the first end. The invention also provides a method for fabricating a conductive antibody probe wafer, wherein a plurality of organic conducting molecules, substantially having an extended and substantially circular chain molecule, are covalently bonded to the basic antibody probe, each of which The conductance antibody probes each of the covalent bonds 201124722 are associated with the second end of the first end of the organic chain of the anchoring molecule corresponding to a certain anchor molecule. [Embodiment] The present invention is to prepare an antibody probe wafer having a covalently bonded anchor molecule, and the substrate of the wafer can use general sand (Si, silicon), silicon nitride (SiO 2, silicon nitride). ), quartz (quartz, yttrium oxide), glass (glass, sulphate) • or mica (including KAlSi04) flat material. The surface of the substrate is coated with a layer of metal, which is preferably applied by, for example, thermal evaporation or e_beam evaporation. Suitable metals include gold (Au (III), silver (Ag), copper (Cu), platinum (Pt), nickel (Ni), zinc (Zn), germanium (Ge), mercury (Hg), palladium (Pd) For example, in the case of the application of the antibody probe wafer, the preferred metal is gold or silver. Gold is generally used because the surface energy of Au(III) in the Au lattice arrangement is the lowest and easier to prepare. If the wafer is used as a sensing wafer, it can be applied to detection such as electrical sensing analysis, for example, as an electrical current sensing chip and an electrical impedance sensing chip. Also as an electrochemical capacitor. The use of (capacitance) and faradaic sensors. In another application, the antibody probe wafer of the present invention is used as a field effect transistor wafer, and can be applied to, for example, a living organism. Detection of Field Effect Transistor Sensing Analysis Method. In another application, the probe wafer of the present invention is used as an oscillating crystal m 11 201124722 sheet, which can be applied to, for example, piezoelectric biosensing analysis. Method detection, for example, quartz vibration micro-plate (quartz cry Stal microbalance. The preferred embodiment of the present invention uses the detection of an electrical material sensing analysis method as an electrical current sensing wafer and an electrical impedance sensing wafer. According to the present invention, the wafer substrate is covered. The metal film layer can be used as a microarray metal substrate arranged in a specific array according to a special design and a specific process. The metal-coated electrode element with the array metal film electrode arrangement can be used for In the biosensing system using the principle of inductance measurement, according to the present invention, a metal film coated on a wafer substrate is chemically modified to have sp3 (gold and sulfur atoms) or sp ( Silver and sulfur atoms are mixed in the form of covalent bonding, thus increasing the stability of the immobilized basic antibody or conducting antibody molecular probe on the surface of the antibody probe wafer substrate. Among the applications, the antibody probe molecule can be an antibody molecule or an antigen-binding ® fragment (Fab). For the application of the wafer, the immobilization of the basic antibody or the conductive antibody probe achieved by the covalent bonding of the present invention can be performed on the wafer in a buffer, and the probe and the sample on the surface of the wafer. An antibody immobilized by covalent bonding on a wafer during the interaction, equilibrium, dissociation, and regeneration steps of the target in the interaction The probe, that is, does not fall off due to the flushing of the strong acid buffer. According to the present invention, the metal film layer coated on the wafer substrate has a new form of chemical bonding, m 12 201124722, which can rapidly immobilize the antibody probe on the surface of the wafer substrate and shorten the fixing of the probe. Time required. The present invention relates to an antibody probe wafer having a covalently bonded organic conductivity (electron conduction) anchor molecule, and the fixed molecular molecule has a cyclic chain molecular structure having an electron transfer capability of a π conjugate system, and is applied to some biochips. Among the uses, the time required for the immobilization of the biomolecule probe can be greatly reduced to three to five minutes from the three to six hours required for the conventional technique to rely on the conventional physical adsorption. • The organic conductivity antibody probe wafer of the present invention is typically applicable to the use of instant sensing wafers. The antibody probe wafer prepared according to the method of the present invention can specifically bind to a specific target functional group in a target molecule, such as a peptide, a protein, a viral surface antigen, or a nucleic acid molecule, to form an affinity probe. An antibody probe having conductivity specificity. Using these conductive specific antibody probes, specific rapid biosensing and measurement can be performed for specific biomedical test samples. According to the preferred embodiment of the present invention, the antibody probe wafer substrate can be a glass wafer used in a microelectronic mechanic system (MEMS) process or a slide used in general optical instruments. The upper cover is vapor deposited with a thin layer of a metal such as gold, silver, copper, uranium, nickel, zinc, bismuth, mercury or IG. Such a metal coated antibody probe wafer substrate can be used as an electrode sheet itself. Since the material of the wafer substrate is a conductive element, it can be applied to a detection instrument generally based on the principle of electrical detection. For example, it is suitable for the detection of electrical, electrochemical electrodes, and transistor electrode analysis formats. [S] 13 201124722 According to the preparation method of the present invention, the organic electrically conductive anchor molecule on the antibody probe wafer substrate bonds the substrate and the probe in a covalently bonded manner and has the ability of electron conduction, thereby increasing the surface of the wafer substrate The stability and electrical conductivity of the immobilized probe. Due to the stability and conductivity of the probe, when the sample is detected, the unbound molecules on the wafer can be washed and removed by buffer to reduce the interference of the impurities on the detection results, so as to improve the probe and the biomedical test sample, Affinity and binding capacity of the target in urine, body fluids, semen, nasal fluid, and laryngeal fluids, and applied to inductance measurements The antibody probe is on the wafer. According to the present invention, in order to bind an immobilized antibody probe on an antibody probe wafer, the selected organic conductivity is first determined to form a covalent bond between the metal on the anchor molecular wafer substrate and directly connected to the metal. The thin film layer is indirectly covalently bonded to the metal thin film layer via a linking molecule. The organic conducting anchor molecule must additionally have a functional group that can undergo a chemical covalent bonding reaction with the antibody probe molecule to be linked. For example, in accordance with several preferred embodiments of the present invention, an organic sulfide (R-SH) can be employed as the primary anchoring molecule on a substrate coated with gold or silver. This is because the sulfur atom easily forms a strong affinity with the transition metal surface such as gold or silver. Wherein, the organic sulfide may be (1) alkyl (tetra) (tetra) sulfide, (2 olig benzene sulphide and (3) oligothiophenes sulfide. In the present invention, the latter two are organic conductance Two preferred preparation examples of anchoring molecules, which are mainly cyclic chain molecular structures having electron conductivity of π-conjugated systems, are mainly oiigopheny1, oligothiophene, oligopyrrole (〇lig〇Pyrr) 〇l) and oligopyrrole 201124722 (oligopyridine) cyclic chain molecular structure. In a preferred embodiment of the invention, firstly, the sulfhydryl sulfide can be placed on the substrate as a binding molecule to indirectly anchor the organic conductance The molecule is bonded to the metal thin film layer. For example, a mercaptoalkylamine having a direct bond thiol group as its main structure has a molecular structure of [HS(CH2)nNH2], wherein n=2 to 16. The sulfur atom of the anchor group forms a sp3-like bond with the surface of the gold metal, while the sulfur atom forms a sp-like bond with the surface of the silver metal. The amine group at the end of the organic molecule (-NH2) Can be retained as a nucleophile For the bonding of the organic conductive anchor molecules, and then the antibody probe-derived bonding. The antibody probe is used to sense the wafer based on the gold or silver metal coating, and the detection is performed because the wafer is solid. It is liquid (such as blood urine, body fluids, saliva, laryngeal fluid, or vaginal fluid, etc.). Therefore, the sensitivity and limit of detection depend on the steric hindrance and degree of freedom of the immobilized probe molecules on the wafer. In addition to maintaining a good degree of freedom between the anchor molecules, the anchor molecule and the probe must also maintain a good degree of freedom to simulate the probe molecule and the analyte in the liquid-liquid phase. The real interaction in the middle. In the case of using the above-mentioned antibody probe-based sensing probe based on the organic conductivity-preserving molecule, the anchor molecule has a cyclic chain molecular structure with electron conduction capability of the π-conjugated system, so The sensitivity and limit of the measured electrical signal (current or impedance ,) depends on the electrical conductivity of the anchoring molecule and the immobilized antibody probe on the metal film. The carbon orbital eight-track domain needs to be re-entered for linearity. 15 201124722 Electron conduction, antibody probe macromolecules, for example, antibody molecule immunoglobulin G molecules (immunoglobulin (3⁄4 IgG) also need to be electronically engineered, soon to be organic The electrically conductive molecule, covalently bonded to the base antibody probe, forms a linear vertical conductive electrode that runs straight from the underlying metal film to the top antigen-binding site of the antibody probe (IgG). For application in the antibody probe sensing chip of the electrical signal. The preparation method of the basic organic light-guided molecular metal substrate capable of bonding the probes of the present invention, the chemical reactions of the first, second and third embodiments respectively show that the preferred embodiment is prepared according to the invention, and the gold-plated (Au) is deposited on the vapor-deposited cloth. On the surface of a metal film substrate of silver (Ag) or copper (Cu), uranium (Pt), and nickel (Ni), the basic organic conducting molecules that can be immobilized are gradually treated by chemical modification. In the preferred embodiment of the present invention relating to the preparation of the first organic conducting anchor molecule, the result of the chemical reaction of Figure 1 is a basic organic conducting wafer substrate 101 which can be coupled to various antibody probes. It can be used to link to the probe probes needed to detect various specific targets for the detection of relevant biological, medical, food or environmental samples. In a preferred embodiment of the present invention, as shown in the chemical reaction of FIG. 1, the organic conductive molecule thiophene sulfide is first fixed on the wafer substrate 110 coated with the gold (Au) metal thin film layer 120. : 5' -(Methyl)thiol-5-aldehyde-2,2'-dithiophene. This can be done by immersing a wafer substrate plated with gold or silver in a 5'-(methyl)thiol-5-aldehyde-2,2'-dithiophene solution at a concentration of about 10 mM, or by immersing it in 201124722. A neutral phosphate buffer solution of about 10 mM 5'-(methyl)thiol-5-acid-2,2'-dithiophene (PBS, pH=7. In 2), the shock is evenly oscillated for about two hours. The gold on the wafer is self-assembled with 5'-(methyl)thiol-5-aldehyde-2,2'-dithiophene. Thereafter, the substrate is cleaned with, for example, ethanol and distilled water to obtain an organic conductive substrate having a surface modification of 5'-(methyl)-5-aldehyde-2,2'-dithiophene. In the present invention, in the second preferred embodiment of the use of organic conductivity-determining anchor molecules, the results of one of the chemical reactions of FIG. 2 are available for the basic organic conductivity wafer substrate 102 which can be coupled to various antibody probes. Link the antibody probes required to detect various specific targets for detection of relevant biomedical, food or environmental samples. In a preferred embodiment of the present invention, as shown in the chemical reaction of FIG. 2, the organic conductive molecular oligophenyl sulfide is fixed on the wafer substrate 110 coated with the gold (Au) metal thin film layer 120. Compound: /> 4'-thiol-4-pylinyl-1,1-diphenyl. This can be achieved by immersing a wafer substrate evaporated with gold or silver at a concentration of about 10 mM; > 4'-thiol-4-keto-1,1-diphenyl in tetrahydrofuran In the solution, the reaction was evenly oscillated for about two hours. The arrangement of gold and -thiol-4-aldehyde-1,1-diphenyl on the wafer is self-assembled. Thereafter, the substrate is washed with, for example, ethanol (ethan〇l) and distilled water to obtain an organic conductive substrate having a surface-modified fluorenyl-thiol-4-aldehyde-U-diphenyl. In the preferred embodiment of the present invention relating to the preparation of the third and fourth organic-conducting anchor molecules, the results of the stepwise chemical reaction of Figure 3 (Fig. 3A and Fig. 3C) are conjugated to each of the 17 201124722 antibody probes. A basic organic conductivity wafer substrate. It can be used to link the antibody probes required to detect various specific targets for the detection of relevant biomedical, food or environmental samples. First, in a preferred embodiment of the present invention, as shown in the chemical reaction of FIG. 3A, an alkyl sulfide is first fixed on the wafer substrate 110 on which the gold or silver metal thin film layer 120 is coated. : 2-thiol-ethanolamine). This can be achieved by immersing a wafer substrate evaporated with gold or silver in a 2-ethylthioethylamine aqueous solution having a concentration of about 1 mM, or immersing in a neutral phosphoric acid containing about 1 mM of 2-thioethylamine. Salt buffer solution (PBS) (pH=7. In 2), the shock is evenly oscillated for about two hours. The arrangement of gold and cysteamine on the wafer is self assembled. Thereafter, the substrate 103 is washed with, for example, ethanol and distilled water to obtain a substrate 103 having a surface modification of 2-thioethylamine. Next, in the preferred embodiment of the present invention relating to the preparation of a third organic-conducting anchor molecule, the wafer chemical reaction of FIG. 3B shows that the substrate obtained by the modification of FIG. 3A is 103, and the first organic conductivity molecule thiophene of the present invention ( Oligothiophene) sulfide derivative: 5"-aldehyde-5-carboxy-2,2',5',2"-trithiophene (5"-aldehyde-5-carboxyl-2,2',5',2" -trithiophene, abbreviated as AC-TTP, having a structure of CHO(C4H2S)2COOH) (100 mM), after shaking for 2 hours, the substrate 104 as shown in Fig. 3B was obtained. The aldehyde group (-CH0) at one end of AC-TTP can be coupled with the primary amine group (-NH2) on the substrate, and the residue at the other end (-COOH) can be reserved as an affinity agent, which is reserved for reaction and then into 201124722. Derivation of biomolecule probes. Thereafter, the wafers were each washed with dichloromethane and ionized water for 10 minutes to remove excess AC-ΤΤΡ molecules on the surface of the wafer. Next, in the preferred embodiment of the present invention for the preparation of the fourth organically used anchoring anchor molecule, the wafer chemical reaction of FIG. 3C shows that the substrate 103 obtained by the modification reaction of FIG. 3 and the second organic conductivity molecular oligobenzene of the present invention. (oligobenzene) sulfide derivative: 4'-wound 4-carboxyl diphenyl (4'aldehyde-4-carboxyl dibenzene, AC-DBZ for short, structure CHO(C6H6)2COOH) aqueous solution (100 mM), Uniform oscillation • After 2 hours of reaction, the substrate 105 as shown in Fig. 3C was obtained. The aldehyde group (-CHO) at one end of AC-DBZ can be coupled with the primary amine group (-NH2) on the substrate, and the thiol group (-COOH) at the other end can be reserved as an affinity agent and retained for the reaction. Derivation of biomolecular probes. Thereafter, the wafers were each washed with a solution of dichlorocarbazine and ionized water for 10 minutes to remove excess AC-DBZ molecules on the surface of the wafer. It is noted that the substrate 1〇3 in Fig. 3A is a semi-finished wafer of the antibody probe wafer. In the organic conductive anchor compound of FIGS. 3B and 3C, it is a derivative compound having a terminal carboxyl group [-COOH], that is, a metal chain coating having a cyclic chain molecular configuration away from the substrate 110. The end of 120 is a carboxyl group. In accordance with the present invention, substrates 104 and 105 are a semi-finished product of an antibody probe wafer. This is due to the carboxyl groups of each anchor molecule on the substrates 104 and 105, and the antibody probes required for any sensing application have not yet been coupled. Such semi-finished substrates 1〇4 and 105 can be stored under appropriate conditions for a long period of time. Typical for at least three months. When required, 201124722, the semi-finished substrates 104 and 105 can be further processed quickly, and any of a variety of antibody probes suitable for covalent bonding with a carboxyl group can be coupled for specific detection applications. According to the present invention, the covalent bonding treatment required for the semi-finished substrate to bind the antibody probe can be extremely fast. Typical treatments can range up to about three minutes of covalent bonding reaction time. Among medical uses, when a new influenza (H1N1) type of disease erupts, the antibody probe design concept according to the present invention, such as the semi-finished substrates 104 and 105 shown in Figures 3B and 3C, can be stocked. The medium is taken out and the probe coupling process is quickly performed to quickly and massively produce an electrical detection antibody wafer for real-time tracking of the disease virus. Example 1: The chemical reaction shown in Fig. 4 shows that it is suitable for linking various types of antibody-detecting basic organic conductive substrate 101, and is capable of linking to various specific antigens for detecting target probes. For the detection of lll (target) in relevant biomedical samples (blood, urine, body fluid, semen, saliva, laryngeal samples). The chemical reaction of Fig. 4 shows that, in an embodiment of the invention, the surface of the base wafer substrate 101 of Fig. 1 is fixed with an organic conductive molecular thiophene sulfide: 5'-(methyl)thiol-5 -Aldehyde-2,2'-mercaptomethyl-5-aldehyde-2,2'-dithiophene (MMAD). The aldehyde group [-CHO] of this organic conductivity molecule can be combined with an antibody probe (probe) The amine group [-NH2] 20 201124722 is subjected to an imine bonding (shiff base) reaction to covalently bond the antibody probe to the organic conductive substrate 101. Thereafter, the composition and the target are formed ( Target) The ability to integrate the ability to measure the wafer substrate 111. a.  In HBS buffer (10 mM HEPES pH 7. 4, 150 mM NaCl, 3. 4mM EDTA, 0. 005% surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P04) A constant flow rate (5 μl/min) was introduced into the flow path, and passed through a wafer substrate 101 containing thiophene sulfide ® MMAD to clean the wafer substrate 105. b.  50 μl of a solution of the antibody probe molecule having an amine group concentration of about 200 μg/ml was injected into the flow channel and passed through the wafer substrate 101 to carry out imine bonding with the aldehyde group [-CHO]. , shiff base) reaction, forming a covalent bond (_CH=N-), covalently bonding the antibody probe to the organic conductivity substrate 101 c.  After the reaction is completed, the balance substrate 111 is washed with a constant flow of HBS buffer to flow the unreacted antibody probe out of the wafer substrate. At this time, the substrate 111 to which the antibody probe-derived compound is covalently bonded to the * terminal shown in Fig. 4 can be obtained. This type of biosensing wafer, which anchors an antibody probe linked to the end of an organic conducting molecule, such as immunoglobulin G, immunoglobulin M, immunoglobulin A (^Immunoglobulin A), immunoglobulin E (Immunoglobulin E), immunoglobulin D (Immunoglobulin D) have a specificity that can be combined with the target to biomedical tests related to post-sputum electrical sensing. 21 201124722 Example 2·· The chemical reaction of Figure 5 shows that the basic organic conductivity substrate 102 can be adapted to link various antibody probes, and can be used to link to various target probes for specific target detection. The preparation method is to perform target detection in relevant biomedical sample shirts, urine, body fluids, semen, saliva, and throat samples. The chemical reaction of Fig. 5 shows that, in one embodiment of the invention, the surface of the base wafer substrate 102 of Fig. 2 is fixed with an organic conductive molecular oligobenzene sulfide: ^>4'-thiol-4 - pf-mercapto-laldehyde dibenzene (MA-DBZ, structure CHO(C6H6)2COOH). The aldehyde group [-CH0] of the organic conductivity molecule can be subjected to an imine bonding (shiff base) reaction with an amine group [-NH2] on the antibody probe (pr〇be) to share the antibody probe. The valence is bonded to the organic conductivity substrate 102. Thereafter, an inductive wafer substrate 112 having the ability to specifically bind to the target is formed. a.  In HBS buffer (10 mM HEPES pH 7. 4, 150 mM NaCl, 3. 4mM EDTA, 0. 005% surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P04) A constant flow rate (5 μl/min) was introduced into the flow path, and passed through the wafer substrate 102 containing the thiophene sulfide MMAD to clean the wafer substrate 102. b.  50 μl of an antibody probe solution having an amine group concentration of about 200 μg/ml was injected into the flow channel and passed through the wafer substrate 102 to carry out m 22 201124722 imine bond bonding with the awake base [-CHO] ( Imine bonding, shiff base) reaction, forming a covalent bond (-CH=N-). The antibody probe is covalently bonded to the organic conductivity substrate 102 c. After the reaction is completed, the equilibrium substrate 112 is washed with a constant flow of HBS buffer to flow unreacted antibody probes out of the wafer substrate. At this time, the substrate 112 having the antibody probe-derived compound covalently bonded to the terminal shown in Fig. 5 was obtained. This type of biosensing wafer, which anchors antibody probes linked to the ends of organic conducting molecules, such as immunoglobulin G, immunoglobulin M, immunoglobulin A (Immunoglobulin A), immunoglobulin E (Immunoglobulin E), and immunoglobulin D (Immunoglobulin D) have the ability to specifically bind to the target, which is beneficial to biomedical tests related to electrical sensing in the future. Example 3: The chemical reaction of Figure 6 shows that the basic organic conductivity substrate 103, which can be adapted to link various antibody probes, can be prepared for attachment to various target probes for specific target detection. The method is for detecting a target in a relevant biomedical sample (blood, urine, body fluid, semen, saliva, laryngeal sample). The chemical reaction of FIG. 6 shows that, in one embodiment of the present invention, the surface of the base wafer substrate 104 of FIG. 3B has an organic conductive oligothiophene sulfonate 5"-aldehyde-5-carboxy-2. 2',5,,2"-trithiophene (5"-aldehyde-5-carboxyl-2,2,,5,,2,,-trithiophene, abbreviation AC-TTP, structure 23 201124722 for CHO(C4H2S)2COOH) The organic conductivity molecule is composed of N-ethyl-N'-(dimethylaminopropyl)carbodiimide (referred to as N-ethyl-N'-(dimethylaminopropyl)carbodiimide in the chemical reaction of Figure 6. E-DAPC), with N-hydroxysuccinimide (HIS), [HO-N-(CO)2(CH2)2)] for the anchoring of carboxylic acid groups of organic conducting molecules [- COOH] performs an amidation reaction to covalently bond an antibody probe to the organic conductivity substrate 104. The molecular structures of HIS and E-DAPC are shown in the reaction of Figure 6, respectively. The HIS/E-DAPC mixture will first activate the carboxylic acid group [-COOH] of the organic conducting molecule, and then combine with an antibody probe having an amine group, etc.: a. In HBS buffer (10 mM HEPES pH 7. 4, 150 mM NaCl, 3. 4mM EDTA, 0. 005% surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P〇4) A constant flow rate (5 μl/min) was introduced into the flow path, and passed through a wafer substrate 104 containing thiophene sulfide AC-TTP to clean the wafer substrate 104. ® b. Will be 50 microliters of 0. 1 M HIS/0. A 4M E-DAPC (1:1) mixed solution was injected into the flow path and passed through the wafer substrate 104 to activate the carboxylic acid group of the AC-TTP. c.  50 μl of a solution of antibody molecules having an amine group concentration of about 200 μg/ml was injected into the flow channel and passed through the wafer substrate 104 to be amidated with the carboxyl group activation group (-CO-O-C4O2H4N) ( Amidation) reaction to form a covalent bond (-CONH.). The antibody probe is covalently bonded to the organic conductivity substrate. d.  After the reaction is completed, the substrate is washed with HBS buffer at a constant current, followed by 5 〇 m 24 201124722 microliters of ethanolamine hydrochloride pH 8. 5) The solution, which is injected into the flow path, flows through the wafer substrate 114 to completely amidate the unreacted activating group (-CO-0-C4〇2H4N). At this time, the substrate 114 to which the terminal molecule is covalently bonded with the antibody molecule-derived compound shown in Fig. 6 can be obtained. This type of biosensing wafer, which anchors an antibody probe linked to the end of an organic conducting molecule, such as immunoglobulin G, immunoglobulin M, immunoglobulin A Φ (^Immunoglobulin Α), immunoglobulin E (Immunoglobulin Ε), immunoglobulin D (Immunoglobulin D) have the ability to specifically bind to the target, which is conducive to biomedical testing related to electrical sensing in the future. Example 4: The chemical reaction of Figure 7 shows that the basic organic conductivity substrate 105, which can be adapted to link various biological probes, can be used to link to various specific target probes for probe detection (probe). A method of preparation for detecting a target in a relevant biomedical sample (blood, urine, body fluid, semen, saliva, laryngeal sample). The chemical reaction of Figure 7 shows that, in a preferred embodiment of the invention, the surface of the basic biochip substrate 105 of Figure 3C has oligo(phenyl) sulfide aldehyde-4-carboxydiphenyl (4'- Aldehyde 4-carboxyl dibenzene, abbreviated as AC-DBZ, is an organic conductivity molecule composed of CHO(C6H6)2COOH). In the chemical reaction of Figure 7, N-diabase succinimide m 25 201124722 N-ethyl- N'-(dimethylaminopropyl)carbodiimide, abbreviated as Ε-DAPC, and N-hydroxysuccinimide, referred to as HSI, [HO-N-(CO)2(CH2)2)] The carboxylic acid group [•COOH] of the conductive molecule AC-DBZ undergoes an amidation reaction to bond the probe probe to the organic conductivity substrate 105. The molecular structures of HSI and E-DAPC are respectively As shown in the reaction scheme of Figure 7, this HSI/E-DAPC mixture will first activate the carboxylic acid group [-COOH] of the organic conductivity molecule AC-DBZ, and then combine with an antibody/probe having an amine group: a .  In HBS buffer (10 mM HEPES pH 7. 4, 150 mM NaCl, 3. 4mM EDTA, 0. 005% surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2PO4) A constant flow rate (5 μl/min) was introduced into the flow path, and passed through a wafer substrate 105 containing oligobenzene sulfide AC-DBZ to clean the wafer substrate 105. b.  Will be 50 microliters of 0. 1 M HIS/0. A 4M E-DAPC (1:1) mixed solution was injected into the flow channel and passed through the wafer substrate 105 to activate the carboxylic acid group of AC-DBZ. c.  50 microliters of a biomolecule solution having an amine group concentration of about 200 micrograms/ml was injected into the flow channel and passed through the wafer substrate 105 to be amidated with the carboxyl group activation group (-CO-O-C4O2H4N) ( Amidation) reaction to form a covalent bond (•CONH-). An antibody probe is covalently bonded to the organic conductivity substrate 105. d.  After the reaction is completed, the substrate is washed with a constant flow of HBS buffer, followed by 50 μl of ethanolamine hydrochloride pH 8. 5) Dissolving m 26 201124722 liquid, injected into the flow path, flowing through the wafer substrate 115 to completely amidate the unreacted activating group (-CO-0-C4〇2H4N). At this time, the substrate 115 to which the antibody probe-derived compound covalently bonded to the terminal shown in Fig. 7 was obtained was obtained. This type of biosensing wafer, which anchors antibody probes linked to the ends of organic conducting molecules, such as immunoglobulin G, immunoglobulin M, immunoglobulin A (immunoglobulin G) Immunoglobulin A), Immunoglobulin E, and Immunoglobulin D have the ability to specifically bind to the target, which is beneficial for biomedical testing related to electrical sensing in the future. Although the present invention has been described above in connection with the preferred embodiments, it is not intended to limit the invention. For example, although the description of the present invention is described in detail by taking an inductive biofilm preparation method as an example, the present invention has an antibody probe wafer having a covalently bonded organic conductivity anchoring compound, and as can be understood, it can also be applied to Others used gold, silver, copper, platinum, nickel, zinc, bismuth, mercury or palladium as treatments for different uses of sensing films on biochips. Applications include, for example, electrical sensing wafers, electrochemical sensing wafers, field effect transistor sensing wafers, piezoelectric sensing wafers, and optical sensing wafers. Therefore, those skilled in the art will be able to make such changes and modifications without departing from the spirit and scope of the invention, and the scope of the present invention is defined by the scope of the appended claims. Example 5: 27 201124722 The chemical reaction of Figure 8 shows one of the embodiments of the invention required to prepare an antibody probe 501 having a surface coated with conduction electrons for related inductivity measurement. Electrical detection of target affinity in medical samples (blood, urine, body fluids, semen, saliva, laryngeal samples). The chemical reaction of Figure 8 shows that, in one embodiment of the present invention, the basic immunoglobulin G antibody molecule 200 of Figure 8 has a surface covalently bonded organic material having the ability to conduct electrons. An organic electron-conducting oligomer 301, for example, a preferred embodiment thereof is 3-(thiophen-2-yl) propanal [(C4H3S)-((C4H3S)-( CH2)-CHO]. The aldehyde group [-CHO] of the organic conductivity oligo molecule can be combined with an antibody probe such as immunoglobulin G, immunoglobulin M, immunoglobulin A, The amine group [-NH2] on immunoglobulin E (Immunoglobulin E) and immunoglobulin D (Immunoglobulin D) undergoes an imine bonding (shiff-base reaction) reaction to modulate the organic conducting oligo molecule 301. Covalently bonded to antibody probe 200. Thereafter, an inductance measuring antibody probe 501 having a specific binding ability to a target is formed. a. With HBS (10 mM HEPES pH 7. 4) Buffer (150 mM NaCl, 3. 4mM EDTA, 0. 005. /. Surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P〇4) A constant flow rate (5 μl/min) was introduced into the flow channel, and the wafer 200 to which the antibody [S] 28 201124722 probe was immobilized was flowed and washed. b.  50 microliters of a 3-(thiophen-2-yl)propanal organic conducting oligo molecule solution 301 having a concentration of about 10 mM was injected into the flow channel and slowly flowed through the antibody probe wafer 200 for 24 hours. It carries out an imine bond reaction with four amine groups on the surface of the antibody molecule, and covalently bonds 3-(thiophen-2-yl)propanal to the antibody probe molecule. c.  After the completion of the reaction, the balanced antibody probe wafer substrate 501 was washed with a constant flow of HBS or PBS buffer to flow unreacted 3-(thiophen-2-yl)propanal oligo molecule 301 out of the wafer substrate. At this time, an antibody probe wafer substrate 501 having a surface covered with 3-(thiophen-2-yl)propanal organic conductance oligo molecule 301 as shown in Fig. 8 was obtained. This type of conductance antibody sensing wafer has the ability to specifically bind to an antigen target, facilitating biomedical testing associated with future electrical sensing. Example 6: $Chemical reaction of Figure 9 is shown as one of the embodiments of the invention required to prepare a conductive electrons antibody probe 502 for performing a related biometric medical sample (blood) , urine, body fluid, semen, saliva, laryngeal fluid) in the target (affinity) electrical detection. The chemical reaction of FIG. 9 shows that, in a preparation example of the present invention, the basic immunoglobulin G antibody molecule 200 of FIG. 9 has a covalently bonded surface covering with conduction electrons. An organic electron-conducting oligomer 302, for example, a preferred embodiment thereof is 29 201124722 3-phenylpropanal [(C6H5)-(CH2)-CHO]. The aldehyde group [-CHO] of the organic conducting oligo molecule can be combined with an antibody probe such as immunoglobulin G, immunoglobulin M, immunoglobulin A, and immunization. The amine group [-NH2] on globulin E (Immunoglobulin E) and immunoglobulin D (Immunoglobulin D) undergoes an imine bonding (shiff-base reaction) reaction to covalently bond the organic conducting oligo molecule 302 After binding to the antibody probe 200, an inductive antibody probe 502 having the ability to specifically bind to the target is formed. a.  With HBS (10 mM HEPES pH 7. 4) Buffer (150 mM NaCl, 3. 4mM EDTA, 0. 005% surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P04) A constant flow rate (5 μl/min) was introduced into the flow path, and the wafer 200 to which the antibody probe was immobilized was flowed and washed. b.  50 μl of a 3-phenylpropionaldehyde organic electro- oligo molecule solution 302 having an aldehyde group of about 10 mM was injected into the flow channel and slowly flowed through the antibody probe wafer 200 for 24 hours to make it and the antibody The amine group on the surface of the molecule undergoes an imine bond-bonding reaction, and 3-phenylpropanal is covalently bonded to the antibody probe molecule. c.  After the reaction is completed, the balanced antibody probe wafer substrate 502 is washed with a constant flow of HBS or PBS buffer to flow unreacted phenylpropanal oligo molecule 302 out of the wafer substrate. At this time, an antibody probe wafer substrate 502 having a surface filled with 3-phenylpropanal organic conductive oligo molecule 302 can be obtained as shown in FIG. 〇 247 201124722 This type of conductivity antibody sensing wafer has an antigen target (target) The ability to combine specificity is conducive to biomedical testing related to electrical sensing in the future. Example 7: The chemical reaction of Figure 10 is not shown as one of the embodiments of the invention required to prepare a conductive electrons antibody probe 503 for performing a related biometric medical sample (blood, Affinity electrical detection in urine, body fluids, semen, saliva, _ throat fluid samples. The chemical reaction of Fig. 10 shows that, in a preparation example of the present invention, the basic immunoglobulin G antibody molecule 200 of Fig. 10 is covalently bonded to the surface and has an organic layer of conduction electrons. An organic electron-conducting oligomer 303, for example, a preferred embodiment thereof is 3-(1-D-pyrrol-2-yl)propanal (3-(l-pyrrol-2-yl) propanal P- (C4H4N)-(CH2)-CHO]. The aldehyde group [-CHO] of this organic conductivity oligo molecule can be combined with an antibody probe such as immunoglobulin (3 (Immunoglobulin G), immunoglobulin M (Immunoglobulin) Μ), Immunoglobulin A, Immunoglobulin E, Immunoglobulin D (Immunoglobulin D), amine group [-NH2] for imine bonding (shiff-base) The reaction is carried out to covalently bond the organic conducting oligo molecule 303 to the antibody probe 200. Thereafter, an inductive antibody probe 5〇3 having a specific binding ability to a target is formed. [S1 31 201124722 a.  With HBS (10 mM HEPES pH 7. 4) Buffer (150 mM NaCl, 3. 4mM EDTA, 0. 005% surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P04) A constant flow rate (5 μl/min) was introduced into the flow path, and the wafer 200 to which the antibody probe was immobilized was flowed and washed. b.  50 μl of a 3-(1-pyridyl-2-yl)propanal organic conductance oligo molecule solution 303 having a concentration of about 10 mM was injected into the channel and slowly flowed through the antibody probe wafer. The imine bond reaction with the four amine groups on the surface of the antibody molecule was carried out for 24 hours, and 3-(1-pyrrol-2-yl)propanal was covalently bonded to the antibody probe molecule. c.  After the reaction is completed, the equilibrium antibody probe wafer substrate 503 is washed with a constant flow of HBS or PBS buffer to flow the unreacted 3-(1-pyridyl-2-yl) agoxylate molecule 303 out of the wafer base fc. An antibody probe wafer substrate 503 having a surface covered with 3-(1-pyridyl-2-yl)propanal organic conductive oligo molecule 303 as shown in Fig. 1A can be obtained. This type of conductance antibody sensing wafer has the ability to bind to antigen target specificity* for biomedical testing associated with future electrical sensing. Example 8: The chemical reaction of Figure 11 is apparent for the preparation of a bioinductor sample (blood, for the purpose of preparing an embodiment of the invention with a conducting electrons antibody probe 504). Affinity electrical detection in urine, body fluids, semen, saliva, and laryngeal fluids. 32 201124722 The chemical reaction of Figure 11 shows that, in one embodiment of the invention, the basic immunoglobulin G antibody molecule 200 of Figure 11 has covalently bonded surface coatings with conducting electrons. An organic electron-conducting oligomer 304, for example, a preferred embodiment thereof is 3-(pyridin-2-yl)propanal [2- (C5H4N)-(CH2)-CHO]. The aldehyde group [-CHO] of the organic conductivity oligo molecule can be combined with an antibody probe (antibody pr〇be> such as immunoglobulin G, immunoglobulin 、, immunoglobulin A (Immunoglobulin Α) The immunoglobulin E (Immunoglobulin E), the amine group [-NH2] on the immunoglobulin D (Immunoglobulin D) undergoes an imine bonding (shiff-base reaction) reaction to expose the organic conducting oligo molecule 304 After covalently bonding to the antibody probe 200, an inductive antibody probe 504 having the ability to specifically bind to a target is formed. a.  With HBS (10 mM HEPES pH 7. 4) Buffer (150 mM NaCl, 3. 4mM EDTA, 0. 0053⁄4 surfactant P20) or PBS (Phosphate buffered saline pH 7. 4) Buffer (137 mM NaCl, 2. 7 mM KC1, 10 mM Na2HP04, 2 mM KH2P〇4) A constant flow rate (5 μl/min) was introduced into the flow path, and the wafer 200 to which the antibody probe was immobilized was flowed and washed. b.  50 μl of a 3-(P-pyrrolidin-2-yl)propanal organic conductance oligo molecule solution 304 having a concentration of about 10 mM was injected into the flow channel and slowly flowed through the antibody probe wafer 2〇〇. For 24 hours, it is reacted with four amine groups on the surface of the antibody molecule by imine bond [S] 33 201124722, and 3-(pyrrole-2-yl)propanal is covalently bonded to the antibody probe molecule. on. c. After the reaction is completed, the balance pit body probe wafer substrate 504 is continuously purged with 甩HBS or PBS buffer to flow unreacted 3-(pyrrole-2-yl)propanal oligo molecule 304 out of the wafer substrate. At this time, an antibody probe wafer substrate 504 having a surface covered with 3-(pyrrolidin-2-yl)propanal organic conductive oligo molecule 304 as shown in Fig. 11 can be obtained. This type of conductance antibody sensing wafer has the ability to specifically bind to an antigen target, facilitating biomedical testing associated with future electrical sensing. Although the present invention has been described above with reference to the drawings in the preferred embodiments, it is not intended to limit the invention. For example, although the description of the present invention is exemplified by a conductive antibody probe wafer, the surface of the present invention is coated with an organic conductive oligo molecule (1) 3-(thiophen-2-yl)propionic acid, (2) 3-phenylpropene. An aldehyde, (3) 3-(1-pyrrol-2-yl)propanoid, (4) 3-(U-pyridin-2-yl)propanal antibody probe, as can be understood, as shown in Figure 12-15, It can also be applied to other derivatives of the above four organic conducting oligo molecules to prepare antibody probes having conduction electrons. Further, although the present invention has been described above with reference to the drawings in the preferred embodiments, it is not intended to limit the invention. As can be appreciated, as shown in Figures 16-21, the same can be applied to other electromechanical oligo-oligo molecules and derivatives thereof having conducting electrons to prepare antibody probes with conduction electrons. Applications include, for example, electrical sensing wafers, electrochemical sensing wafers, field effect transistor sensing wafers, piezoelectric sensing wafers, and the like. Therefore, any such modifications and variations can be made by those skilled in the art without departing from the spirit and scope of the invention, and the scope of the invention is defined by the scope of the appended claims. quasi.

[si 35 201124722 [Simplified illustration of the drawings] The chemical reactions of Figures 1, 2 and 3 respectively show the use of chemical modification on the surface of a substrate coated with gold or silver coated according to a preferred preparation example of the present invention (chemieal • After the treatment, a cyclic organic conductive oligothiophene and an oligobenzene molecule of a specific length are immobilized for attachment to a basic antibody probe wafer substrate of an antibody to be used. • The chemical reaction in Figure 1 shows that on the wafer substrate coated with the metal film of Au (metal) gold, the chemical modification is carried out by covalent bonding with sp3 (gold and sulfur atoms). A molecular oligothiophene sulfide: for example, 5'-(methyl)thiol-5-aldehyde-2-,2'-dithiophene, attached to a metal film. The chemical reaction of Figure 2 shows that on the wafer substrate coated with the vapor deposited gold metal thin film layer, the chemical modification is carried out by covalent bonding of sp3 (gold and sulfur atom) mixed domains to the organic conductive molecular oligophenyl (oligophenyl). Sulfide: for example; > 4'-thiol-4-aldehyde group φ-1,1-diphenyl. , fixed to the metal film. The chemical reaction of Fig. 3A shows that an alkyl sulfide: thiol-ethanolamine is immobilized on a wafer substrate on which a gold or silver metal thin film layer is coated. The wafer chemical reaction of Fig. 3B shows that on the surface of the antibody probe wafer substrate of Fig. 3A, the organic conductive molecule thiophene (oHgothiophene) sulfide derivative: 5"-awake-5-carboxy-2, 2', 5', 2"-trithiophene (5"-aldehyde-5-carboxyl-2,2',5',2"-trithiophene), the awake group (-CHO) at one end can be carried out with the primary amine group (-NH2) on the substrate. The coupling reaction, the carboxyl group 36 201124722 base (-COOH) at the other end can be reserved as a affinity agent for the derivatization of the antibody molecule probe after the reaction. The wafer chemical reaction of Figure 3C shows the use of the organic conductivity molecular oligobenzene sulfide derivative on the surface of the antibody probe wafer substrate of Figure 3A: 4'-aldehyde-4-carboxydiphenyl (4'-aldehyde-4-carboxyl) Dibenzene), one end of the wake-up (-CHO) can be coupled with the primary amine group (-NH2) on the substrate, the other end of the carboxyl group (-COOH) can be reserved as a affinitive agent, reserved for the reaction after the antibody Derivatization of the probe. The chemical reaction of Fig. 4 shows that an organic conductive molecular oligothiophene sulfide derivative is immobilized on the surface of the basic antibody wafer substrate of Fig. 1. The aldehyde group [-CHO] of the organic conductivity molecule can be imine-bonded with the amine group [-NH2] on the antibody probe to covalently bond the antibody probe to the organic On the conductivity substrate. The chemical reaction of Fig. 5 shows that an organic conductive molecular oligophenyl sulfide derivative is immobilized on the surface of the basic antibody wafer substrate of Fig. 2. The aldehyde group [-CHO] of the organic conductivity molecule can be subjected to an imine bonding reaction with an amine group [-NH2] on an antibody probe (Ab probe) to covalently bond the antibody probe to On the organic conductivity substrate. The chemical reaction of Fig. 6 shows that an organic conductive molecular oligothiophene sulfide derivative is immobilized on the surface of the basic antibody wafer substrate of Fig. 3B. The carboxylic acid group [-COOH] of the organic conductivity molecule can be subjected to an amidation reaction with an amine group [-Nm] on an antibody probe to covalently bond the antibody probe. To have 37 201124722 on the electromechanical guide substrate. The chemical reaction of Fig. 7 shows that an organic conductive molecular oligobenzene sulfide derivative is immobilized on the surface of the basic antibody wafer substrate of Fig. 3C. The carboxylic acid group [-COOH] of the organic conductivity molecule can be subjected to an amidation reaction with an amine group [-NH2] on a basic antibody probe to covalently bind a basic antibody probe (probe) Bonded to the organic conductivity substrate. The chemical reaction of Figure 8 shows that the conductive antibody probe flows through the surface of the basic antibody probe wafer on the surface of the substrate, through an organic electron-conducting moleecule 3- (thiophene-2-) having conducting electrons. 3-(thiophen-2-yl) propanal and derivatives The awake base of the organic conducting molecule [-CH0] can be imidized with the amine group [-NH2] on the antibody probe. (imine bonding) covalent bonding reaction to coat the organic conductive molecules onto the antibody probe. The chemical reaction in Figure 9 shows that the conductive antibody probe flows through the surface of the basic antibody probe wafer substrate and has conductive electrons. The organic conductive molecule 3-phenylpropanal and derivatives. The organically conductive molecule of the awake base [-CH0] can be imidized with the amine group [-NH2] on the antibody probe. (imine bonding) covalent bonding reaction to coat the organic conductive molecules onto the basic antibody probe. The chemical reaction in Figure 10 shows the conductivity antibody probe flowing through the surface of the basic antibody probe wafer substrate. Organic Conductance Molecules 3- (1-pyrrol-2-yl)propanal and its derivatives (3-(l-pyir〇l-2-yl)propanal and derivatives). The acid group [-CH0] of this organic conductivity molecule can be combined with an antibody probe. The amine group [-NH2] is subjected to a covalent bonding reaction of imine bond (imine 38 201124722 bonding) to coat the organic conductive molecule onto the antibody probe. The chemical reaction of Fig. 11 shows that the basic antibody probe The surface of the wafer substrate flows through 3-(pyridin-2-yl)propanal and its derivatives (3-(pyridin-2-yl)propanal and derivatives), which have conductive electrons. The base [-CH0] can be subjected to a covalent bonding reaction with an amine group [_ leg 2] on the antibody probe to coat the organic conductive molecule onto the antibody probe. They are respectively applicable to the structural formula of the organic conducting molecule and its derivative which can be used for the above-mentioned electrical conductivity antibody probe of the present invention to prepare a conducting antibody probe having conduction electrons, and the chemical formulas thereof are: (1) . 3-( Thiophen-2-yl) propylene and its derivatives 3-(thiophen-2-yl)propanal and derivatives 〇 see Figure 12. (2). 3-Phenylpropionic acid and its derivatives Bio 3-phenylpropanal and derivatives. See Figure 13. (3) 3-(l-D than alk-2-yl) adenine and its derivatives 3-(l-pyrrol-2-yl)propanal and lu derivatives. See Figure 14. (4). 3-(p-pyridyl-2-yl) azepine and its derivative 3-(pyridin-2-yl)propanal and derivatives 〇 Figure 15〇(5) .cis-4-phenyl -3- Ding waking and its derivatives (6)-4_phenylbut-3-enal and derivatives. See Figure 16. (6) cis-4-(pyridin-2-yl)-3-butenal and its derivatives (£)-4, (pyridin-2-yl)but-3-enal and derivatives. See Figure 17. 39 201124722 (7). cis-seven-4,6-di-dialdehyde and its derivatives (6)-hepta-4,6-dienal and derivatives. See Figure 18. (8). Trans-seven-4,6-two awake and its derivatives (Z)-hepta-4,6-dienal and derivatives. See Figure 19. (9) . cis-4-(cyclohex-2-enyl)butanal and its derivatives (5)-4-(cyclohex-2-enylidene) butanal and derivatives. See Figure 20. (10) 2-(cyclohydrocarbon-2-yl) acetonide and its derivatives 2-(naphthalen-2-yl)acetaldehyde and derivatives. See Figure 21. It should be noted that, for the novelty of the present invention, the various preferred preparation examples described above in conjunction with the drawings only describe in detail the several types of electron conduction performed by the present invention in a sensing wafer system. One of the possible practices, that is, the organic conductivity molecule is also used as a linker for the antibody counter electrode, ie, as depicted in Figures 4, 5, 6 and 7. However, as will be appreciated by those skilled in the art, other arrangements of organic conductance molecules (conductive attractants) in the system of the present invention are equally possible and feasible. For example, a slightly different arrangement involves the use of organic conductivity molecules to electronically engineer antibodies, i.e., those depicted in Figures 8, 9, 10 and 11. The organic conducting molecule is added to the buffer flowing through the surface of the sensing wafer, or an organic conducting molecule is added to the sample of the sample. Or even any combination of these arrangements, as it has been experimentally proven, is a viable practice. 201124722 [Main component symbol description]

Claims (1)

201124722 VII. Patent application scope: 1. A method for fabricating a conductive antibody probe wafer, comprising: a substrate having a metal film layer deposited on a surface of the substrate; and a plurality of organic conductance anchor molecules having substantially Extending and substantially in the form of a cyclic chain molecule, each of the organic electrically conductive anchor molecules is directly covalently bonded to the metal thin film layer by one of the first ends of the cyclic chain molecular structure, or indirectly through the linker The price is bonded to the metal thin film layer; and a plurality of basic antibody probes, each of the basic antibody probes are covalently bonded to the organic conductive anchor molecules corresponding to certain anchor molecules against the ring a second end of the first end of the chain molecular structure; and a plurality of electrically conductive antibody probes, wherein the plurality of organic conducting molecules, the molecules having substantially extended and substantially cyclic chains, are covalently bonded On the basic antibody probe, each of the conductive antibody probes is covalently bonded to the organic conductive derivative anchors* corresponding to a certain anchor molecule against the molecular structure of the cyclic chain One end of the second end. 2. The method for preparing a basic antibody probe wafer according to claim 1, wherein the anchor molecule is covalently bonded to the metal thin film layer at the first end, and the cyclic molecular structure is an organic oligothiophene. (oligo-thiophene) sulfide having the structure [HS-(CH2)n-(C4H2SHC4H2S)m-CHO], wherein n = 0 to 12, m = 0 to 6. 3. For example, the anchor molecule [S] 42 201124722 [HS-(CH2)n-(C4H2S)-(C4H2S)m-Xl· can be directly used with the antibody probe at its second end. The valence chain, the second terminal functional group -X is an aldehyde group [-CHO], a carboxylic acid group [COOH], a disulfide bond [_S-S-], an amine group [m], or a thiol group [- SH] 〇4. The basic antibody probe wafer maker according to item 1 of the patent application scope & wherein the anchor molecule can be directly bonded to the metal thin film layer at the first end, and the cyclic chain molecular structure thereof It is an organo oligo-benzene sulfide with a structure of [HS-(CH2)n-(C6H4HC6H4)m-CHO], where n = 0~12, m = 0~6. 5. The anchor molecule according to item 4 of the patent application can be directly covalently bonded to the antibody probe at its second end, and the second terminal functional group -X is an aldehyde group [-CHO], a carboxylic acid group. [-COOH], disulfide bond [-SS-], amine group [-NH2], or thiol group [-SH]. 6. The method of fabricating a basic antibody probe wafer according to claim 1, wherein the anchoring molecule is indirectly bonded to the metal thin film layer through a linker, wherein the linker is an organoalkyl sulfide, The structure is [HS-(CH2)n-NH2], wherein n = 1~7 7. The basic antibody probe wafer preparation method according to claim 1, wherein the first end of the anchor molecule can be indirectly transmitted through the linker Covalently bonded to the metal thin film layer, wherein the anchoring molecule is an organic oligothiophene sulfide derivative having the structure [CHCKCHA-CC^SMCftSVCOOH], wherein n = 0~12, m = 0~6. 8. For example, the anchor molecule [CHO-(CH2)n-(C4H2S)-(C4H2S)mX] of claim 7 may be covalently linked to the base antibody 43 201124722 probe at the second end thereof. The second terminal functional group -X is aldehyde group [-CHO], carboxylic acid group [-COOH], disulfide bond [-SS-], amine group [-NH2], or thiol group [-SH] 〇9 The basic antibody probe wafer maker according to claim 1 of the patent application, wherein the first end of the anchor molecule is covalently bonded to the metal thin film layer through a linker, wherein the anchor molecule is An organic oligobenzene sulfide derivative having the structure [CHO-(CH2)n-(C6H4)-(C6H4)m-COOH], wherein η = 0 〜12, m = 0 〜6. 10. For example, the anchor molecule of the ninth application patent [CHO-(CH2)n-(C6H4)_(C6H4)mX], wherein the second end thereof can be covalently linked with the basic antibody probe, the first The terminal functional group -X is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [·ΝΗ2], or a thiol group. 11. The method for fabricating a conductive antibody probe wafer according to claim 1, wherein a plurality of organic conducting molecules are covalently bonded to the basic antibody probe, wherein the organic conducting molecular system is organic 3- (thiophen-2-yl)propanal and its derivative 3-(thiophen-2-yl)propanal and derivatives, the structure is [H-(C4H2S)n-(CH2)mX], where η = 1,2,3 , 4, m = 0 to 12. 12. The organic conductivity molecule [H-(C4H2S)n-(CH2)mX] as claimed in claim 11 wherein one end functional group -X can be covalently bonded to a basic antibody probe, the functional group -X It is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [-NH2], or a thiol group [-SH]. 13. The method of fabricating a conductive antibody probe wafer according to claim 1, wherein a plurality of organic conducting molecules are covalently bonded to the basic antibody probe, [S] 44 201124722 wherein the organic conducting molecule The structure is 3-phenylpropanal and derivatives, and the structure is [H-(C6H4)n-(CH2)mX], where η = 1,2,3,4, m = 0 to 12. 14. The organic conductivity molecule [H-(C6H4)n-(CH2)mX] as claimed in claim 13 wherein one end functional group -X can be covalently bonded to a basic antibody probe, the functional group -X It is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [-NH2], or a thiol group [-SH]. 15. The method of fabricating a conducting antibody probe wafer according to claim 1, wherein a plurality of organic conducting molecules are covalently bonded to the basic antibody probe, wherein the organic conducting molecular system is organic 3- (1-(l-pyrrol-2-yl) propanal and derivatives), the structure is [H-(C4H3N)n-(CH2)mX], wherein η = 1,2,3,4, m = 0 to 12. 16. The organic conductivity molecule [H-(C4H3N)n-(CH2)mX} of claim 15 wherein the one-end functional group -X can be covalently bonded to the antibody probe, the functional group -X is Aldehyde [-CHO], carboxylic acid group [-COOH], disulfide bond [-SS-], amine group [-NH2], or thiol group [-SH] 〇 17. As in the scope of claim 1 A method for fabricating a conductive antibody probe wafer, wherein a plurality of organic conductivity molecules are covalently bonded to a basic antibody probe, wherein the organic conductivity molecule is an organic 3-(pyrrole-2-yl)propanal And its derivatives (3-(pyridin-2-yl)propanal and derivatives), the structure is [H-(C5H3N)n-(CH2)mX], where η = 1,2,3,4, m = 0 〜 12. [SI 45 201124722 18. The organic conductivity molecule [H-(C5H3N)n-(CH2)mX} as claimed in claim 17 wherein the one-end functional group -X can be covalently bonded to the antibody probe, the functional group -X is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [m], or a thiol group [_SH] 〇 19. The method for producing a conductive probe probe wafer, wherein a plurality of organic conductive molecules are covalently bonded to a basic antibody probe, wherein the organic conductive molecular system is (1) cis-4-phenyl 3-butenal and its derivative organisms (£)-4-phenylbut-3-enal and derivatives [H-(C6H4-C2H2)n-(CH2)mX], (2) cis-4-(pyridine -2·yl)-3-butenal and its derivatives (&)-4-(pyridin-2-yl)but-3-enal and derivatives [H-(C5H3N_C2H2)n-(CH2)mX], (3) cis-seven-4,6-dienal and its derivatives (£)-hepta-4,6-dienal and derivatives [H-(C2H2_C2H2)n-(CH2)mX], (4) trans-seven- 4,6-two awake and its derivatives (2)-1^1&-4,6-(^1^1&11(1(161'^1^5 [H-(C2H2-C2H2)n- (CH2)mX], (5) cis-4-(cyclohex-2-enyl)butanal and鲁 生物 (£)-4-(cyclohex-2-enylidene) butanal and derivatives [H-(C6H7=CH)n-(CH2)mX], (6) 2-(cycloalkyl-2-yl)acetaldehyde And its derivative 2-(naphthalen-2-yl)acetaldehyde and derivatives, [H-(C4H4_C6H3)n-(CH2)mX], where η = 1,2,3,4, m = 0~12. For example, the organic conductivity molecules in the 19th article of the patent application are [H-(C6H4-C2H2)n-(CH2)mX], [H-(C5H3N-C2H2)n-(CH2)mX], [H-(C2H2) -C2H2)n-(CH2)mX] , [H-(C2H2-C2H2)n-(CH2)mX], [S) 46 201124722 [H-(C6H7=CH)n-(CH2)mX] and [H -(C4H4-C6H3)n-(CH2)mX], wherein one end functional group -x can be covalently linked to an antibody probe, the functional group -x is an aldehyde group [-CHO], a carboxylic acid group [- COOH], disulfide bond [-SS-], amine group [-NH2], or thiol group [-SH]. The method of producing an antibody probe wafer according to any one of claims 1 to 10, wherein the metal thin film layer is a gold, silver, copper, platinum, rhodium, zinc, ruthenium, mercury or palladium film layer. The method of producing an antibody probe wafer according to any one of claims 1 to 20, wherein the metal thin film layer is an array metal thin film layer. 23. A probeless semi-finished wafer substrate of a conductive antibody probe wafer can be pre-stored to couple the wafer for probe sensing when needed, the probeless semi-finished wafer substrate comprising: a substrate having a metal film layer on a surface thereof, wherein the metal film layer is a gold, silver, copper, platinum, nickel, zinc, lanthanum, mercury, palladium film layer; and the plurality of substantially extending and substantially The anchoring molecule of the cyclic chain molecular structure, the first end of one of the respective cyclic chain molecular structures, is directly covalently bonded to the metal thin film layer, or indirectly covalently linked to the metal thin film layer through the linker. And a basic antibody probe covalently bonded to each of the anchor molecules corresponding to the second end of the first end. The basic antibody probe can be covalently bonded to the need to form the basic antibody probe wafer, and a plurality of electrically conductive antibody probes, wherein the plurality of organic conductivity molecules are substantially 47 201124722 a molecule extending and substantially in the form of a cyclic chain, covalently bonded to the basic antibody probe, each of the conductive antibody probes being covalently bonded to the corresponding organic anchoring anchor molecule A second end of the first end of the cyclic chain molecular structure. [ 48