TWI475227B - Antibody probe chip with covalent bonding electron-conducting molecule - Google Patents

Description

Antibody probe wafer with covalently bonded electron conducting molecules

The invention relates to an electron-conducting antibody probe chip, in particular to a basic antibody probe chip, which is covalently bonded to an organic conductive anchor molecule on a metal substrate. In addition to the electron-conducting anchoring molecule, the basic antibody probe is also covalently bonded to the organic-conducting molecule. Therefore, the stability of the immobilization probe on the surface of the wafer substrate and the ability of electron conduction can be increased, and the antibody probe wafer for measuring the sample inductance can be quickly and sensitively performed.

An antibody probe chip 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 which the detection is made may be a biomolecule, virus, bacterium or cell present in the sample. 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, immunoglobulin. Protein E (Immunoglobulin E), Immunoglobulin D or its antigen-binding fragments, or other viruses that can react or catalyze with the target to be tested, Cell or tissue surface biomolecules. Such a substrate is coupled to an immobilized antibody or a fragment thereof as a probe for affinity with a biomolecule (ie, a target to be analyzed) in the biomedical sample. A basic data diagnosis and experimental parameters can be provided by detecting the binding concentration or binding constant of the interaction between the two. The high affinity, high binding capacity and biostability between the two are very important features in the detection of biochips.

There are a variety of bioassays that require the immobilization of biomolecules such as proteins directly onto substrates. For example, western blotting adsorbs proteins on polyvinylidene fluoride (PVDF) or nitrocellulose membrane substrates as probes, and can interact with surface functional groups on antibodies or antigenic proteins. Interaction to measure whether the sample contains such protein. In addition, another example is an enzyme-linked immuno-sorbent assay (ELISA), which adsorbs a specific antibody (or antigen) on a polystyrene substrate and exposes the substrate to In the sample. If the sample contains a specific antigen (or antibody) with high affinity for the antibodies (or antigens), it will be detected by the light output.

For example, Rampal et al., US Pat. No. 6,013,789, "Covalent attachment of biomolecules to derivatized polypropylene supports", discloses that a polypropylene film is first subjected to amination, followed by a polynucleotide containing a phosphorimidazolide at its end. The reaction, thus, the polynucleic acid can be covalently linked to the polypropylene via a phosphoramidate linkage.

In addition, by using self-assembly of molecules on the surface of indium tin oxide (ITO) to form self-assembled monolayers, a new interface can also be provided, which can be used to explore the specificity of certain biomolecules. Redox reactions. The organic ruthenium may be reacted with a substrate having a hydroxyl group on the surface (for example, ruthenium dioxide) to form a single-layer or two-layer organic ruthenium film. For example, Chrisy et al., in the "Selective attachment of biomolecules to patterned self-assembled surfaces" case of US Pat. No. 5,688,642, proposes an organic ruthenium compound having two reactive groups, utilizing an active group at one end and a hydroxyl group on the surface of the substrate. A linkage is generated, and the biomolecule is linked by an active group at the other end to allow the biomolecule to 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 to electrical students. Testing 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 cleaning procedure is continuously performed several times. After cleaning, the blocking reagent is added and allowed to stand for a long time (usually about three to four hours) before the wafer can be used as a biomolecular probe wafer.

Therefore, there is a need to provide a biomolecular probe chip, particularly an antibody probe chip, having covalently bonded electron-conducting anchoring molecules, or antibodies thereon. The organic-conducting molecules are coated, and the wafers have the ability to conduct electrons with conduction electrons. The preparation is simple and fast, and has high stability and specificity in storage and use, and can be fast and accurate. Perform an electrical test of the sample.

In order to achieve the foregoing and other objects, the present invention provides a basic antibody probe wafer having a covalently bonded organic electrically conductive anchor molecule, comprising: a substrate having a metal thin film layer on one surface thereof; and a plurality of The organic conductance anchor molecule has a cyclic chain molecular structure with an electron transfer capability of a π-conjugated system, mainly oligophenyl, oligothiophene, oligopyrrol and oligopyridine ring. a chain-like molecular structure , each of which is an organic electrically conductive anchor molecule, each of which is directly covalently bonded to the metal thin film layer at one of the first ends of the cyclic chain molecular structure, or indirectly covalently bonded to the bonded molecule On the metal thin film layer; and a plurality of probe molecules, each of the basic antibody probes are covalently bonded to the anchor molecules, and the corresponding anchor molecules are opposite to the first end of the cyclic chain molecular structure One of the second ends. The invention also provides 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, and each of the conducting antibodies Each of the 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 of an antibody probe wafer, which can be pre-stored to couple the antibody probe wafer to detect probe molecules required for sensing when needed, and the probeless semi-finished wafer substrate includes a substrate having a metal thin film layer on a surface thereof; and a plurality of organic conductive anchor molecules, and a cyclic chain molecular structure substantially having an electron transfer capability of a π-conjugated system, each of the anchor molecules The first end of one of the cyclic chain molecules is directly covalently bonded to the metal thin film layer, or indirectly covalently bonded to the metal thin film layer through a linking molecule; and it opposes the circular chain molecular structure A second end of the first end can be covalently bonded to the probe molecule as needed to form the molecular probe wafer. The invention also provides 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, and each of the conducting antibodies Each of the probes is covalently bonded to the second end of the first end of the organic chain of the molecular structure.

The present invention is covalently bonded wafer probe anchor given antibody molecule, which substrate wafers (Substrate) using a general silicon-based (Si, silicon), silicon nitride (SiO _2, silicon nitride), quartz (Quartz , yttrium oxide component), glass (glass, silicate) or mica (containing KAlSiO ₄ ) 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 use of the antibody probe wafer, the preferred metal is gold or silver. Gold is generally used because Au (III) has the lowest surface energy and is easy to prepare in the Au lattice arrangement.

The antibody probe wafer of the present invention, if used as a sensing wafer, can be applied to detection such as electrical sensing analysis, for example, as an electrical current sensing chip and an electrical impedance sensing wafer. . Also used as an electrochemical capacitor and a faradaic sensor. In another use, the antibody probe wafer of the present invention is used as a field effect transistor wafer and can be applied to detection such as biofield effect transistor sensing analysis. In another use, the probe wafer of the present invention is used as an oscillating wafer, and can be applied to detection such as piezoelectric biosensing analysis methods, for example, quartz crystal microbalance (quartz crystal microbalance) ). The preferred embodiment of the present invention uses the detection of an electrical material sensing analysis method for use as an electrical current sensing wafer and an electrical impedance sensing wafer.

According to the present invention, the metal thin film layer coated on the wafer substrate can be a microarray metal substrate arranged in a specific array after being specially designed and processed by a specific process. The metal-clad electrical sensing component with the array metal film electrode arrangement can be used on a biosensing system applying the principle of inductance measurement.

According to the present invention, a metal film of a metal film coated on a wafer substrate is chemically modified by using a sp3 (gold and sulfur atom) or sp (silver and sulfur atom) mixed orbital covalent bond. The form of covalent bonding thus increases the stability of the immobilized basic antibody or conductance antibody molecular probe on the surface of the antibody probe wafer substrate. In the application of the biochip, the antibody probe molecule may be an antibody molecule or an active-binding fragment (Fab). In the application of the biochip, the basic antibody or the conductive antibody probe obtained by the covalent bonding of the present invention is immobilized, and the wafer can be processed in a buffer, and the probe and the probe on the surface of the wafer are used. The association between the target in the sample, the equilibrium, the dissociation, and the regeneration step, the wafer is immobilized by covalent bonding. The antibody 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 is chemically bonded to provide a new form for rapidly immobilizing the antibody probe on the surface of the wafer substrate, thereby shortening the time required for immobilization of the probe. . The present invention relates to an antibody probe wafer having a covalently bonded organic conductance (electron conduction) anchor molecule, and the anchor molecule has a cyclic chain molecular structure having an electron transfer capability of a π-conjugated system, and is applied to some biochips. Among the uses, the time required for 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 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. With these conductive specific antibody probes, specific rapid biosensing and measurement can be performed for specific biomedical test samples.

According to a preferred embodiment of the present invention, the antibody probe wafer substrate may be a glass wafer used in a microelectronic mechanical system (MEMS) process or a glass slide used in general optical instruments. The coating is vapor deposited with a thin layer of a metal such as gold, silver, copper, platinum, nickel, zinc, bismuth, mercury or palladium. 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.

According to the method of the present invention, the organic electrically conductive anchor molecules on the antibody probe wafer substrate link the substrate and the probe in a covalently bonded manner and have the ability of electron conduction, thereby increasing the stability of the immobilized probe on the surface of the wafer substrate. Sexual and electrical conductivity. Due to the stability and electrical conductivity of the probe, the unbound molecules on the wafer can be removed by buffer washing during sample detection, reducing the interference of impurities on the detection results, and improving the probe and biomedical test samples (blood, Affinity and binding capacity between the target in urine, body fluids, semen, nasal fluid, and laryngeal fluids, and applied to antibodies related to inductance measurement On the probe 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 electrically conductive 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 wafer 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. Among them, the organic sulfide may be (1) an alkyl sulfide, (2) an oligobenzene sulfide, and (3) an oligothiophenes sulfide. In the present invention, both the guide and the given two kinds of organic molecules of the preferred embodiment of the anchor, which is a main chain molecular structure having a cyclic π system of the electronic conductivity of the conjugated oligonucleotides predominantly benzene (oligophenyl), Oligothiophene, oligopyrrol and oligopyridine cyclic chain molecular structure.

In one preferred embodiment, the present invention can be first placed as alkyl sulfides coupling organic molecules indirectly guide the anchoring of the molecules attached to the metal thin film layer on the substrate. For example, a mercaptoalkylamine having a linear alkyl group as its main component has a molecular structure of [HS(CH ₂ ) _n NH ₂ ], wherein n = 2 to 16. The sulfur atom, which acts as an anchor group, forms a sp ³ -like bonding morphology with the surface of the gold metal. The sulfur atom forms a sp-like bond with the surface of the silver metal. The amine group (-NH ₂ ) at the end of the organic molecule can be retained as a nucleophilic agent to facilitate the bonding of the organic conductive anchor molecule, thereby performing antibody probe-derived bonding.

The wafer is sensed by the aforementioned antibody probe based on gold or silver metal coating. When the detection is performed, the object to be detected is liquid (such as blood urine, body fluid, saliva, throat fluid, or vaginal fluid). Etc.) Therefore, the sensitivity and limits 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 anchoring molecules, a good degree of freedom must be maintained between the anchoring molecule and the probe to simulate the probe molecule and the analyte in the liquid-liquid phase. Real interaction.

In addition, when the above-mentioned antibody probe-based sensing chip based on the organic conductive anchor molecule is used for detecting, the anchor molecule has a cyclic chain molecular structure having a π-conjugated system electron conduction capability, and thus the detected electrical signal (current) The impedance and the sensitivity are based on the electrical conductivity of the anchoring molecules and the immobilized antibody probes on the metal film. Each anchoring elements P _z orbitals of carbon atoms required for linear electron conduction overlap, the giant molecule antibody probes, e.g., antibody molecule immunoglobulin G molecule (immunoglobulin G, IgG) should also be electronically engineered, i.e. organic conductance The 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.

Preparation of basic organic conductivity molecular metal substrate capable of coupling antibody probe

The chemical reactions of Figures 1, 2 and 3 respectively show gold (Au), silver (Ag) or copper (Cu), platinum (Pt), nickel (Ni) in the vapor deposition coating according to several preferred embodiments of the present invention. On the surface of the metal thin film substrate, the basic organic conductive molecules that can be immobilized are gradually treated by chemical modification.

In the preferred embodiment of the present invention relating to the use of an organic electrically conductive anchoring molecule, the result of the chemical reaction of Figure 1 is a substantially organic conducting wafer substrate 101 to which various antibody probes can be coupled. It can be used to link the antibody probes required to detect various specific targets for the detection of related 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 immobilized 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 achieved by immersing a wafer substrate evaporated with gold or silver in an aqueous solution of 5'-(methyl)thiol-5-aldehyde-2,2'-dithiophene at a concentration of about 10 mM, or immersing in about 10 In a neutral phosphate buffer solution (PBS, pH=7.2) of mM 5'-(methyl)thiol-5-aldehyde-2,2'-dithiophene, the reaction was uniformly shaken 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 washed 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 preferred embodiment of the present invention relating to the second embodiment of the use of an organic conductivity anchoring molecule, the result of one of the chemical reactions of FIG. 2 is a basic organic conductivity wafer substrate 102 that can be coupled to various antibody probes. It can be used to link probe probes for the detection of various specific targets for the 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: p -4'-thiol-4-aldehyde-1,1-diphenyl. This can be carried out by immersing a wafer substrate evaporated with gold or silver in a tetrahydrofuran solution of p -4'-thiol-4-aldehyde-1,1-diphenyl at a concentration of about 10 mM. , evenly oscillate for about two hours. The gold on the wafer is self-assembled with p -4'-thiol-4-aldehyde-1,1-diphenyl. Thereafter, the substrate is washed with, for example, ethanol and distilled water to obtain an organic conductive substrate having a surface modification of p- 4-thiol-4-aldehyde-1,1-diphenyl.

In the third and fourth preferred embodiments of the present invention, the results of the stepwise chemical reaction of Figure 3 (Fig. 3B and Fig. 3C) are the basic organic conductances of the various antibody probes that can be coupled. Wafer substrate. It can be used to link probe probes for the detection of 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 done by immersing a wafer substrate evaporated with gold or silver in an aqueous solution of 2-thioethylamine having a concentration of about 100 mM, or immersing in a neutral phosphate buffer solution (PBS containing about 100 mM of 2-thioethylamine). (pH = 7.2), the homogeneous shock reaction is 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 a preferred embodiment of the present invention relating to the use of an organic electrically conductive anchoring molecule, the wafer chemical reaction of FIG. 3B shows that the substrate 103 obtained by the modification of FIG. 3A, and the first organic conducting molecule oligothiophene of the present invention. Sulfide derivative: 5"-aldehyde-5-carboxy-2,2',5',2"-trithiophene (5"-aldehyde-5-carboxyl-2,2',5',2"-trithiophene Referring to AC-TTP for short, the structure is CHO(C ₄ H ₂ S) ₂ COOH) aqueous solution (100 mM), and after shaking for 2 hours, the substrate 104 as shown in Fig. 3B can be obtained. The aldehyde group (-CHO) at one end of the AC-TTP can be coupled with the primary amine group (-NH ₂ ) on the substrate, and the carboxyl group (-COOH) at the other end can be reserved as a affinity agent, and retained for the reaction. Derivation of biomolecular probes. Thereafter, the wafers were each washed with dichloromethane and ionized water for 10 minutes to remove excess AC-TTP molecules on the wafer surface.

Next, in the fourth embodiment of the present invention relating to the use of an organic conductivity anchoring molecule, the wafer chemical reaction of FIG. 3C shows that the substrate 103 obtained by the modification reaction of FIG. 3A and the second organic conductivity molecular oligobenzene of the present invention. ) sulfide derivative: 4 '- biphenyl-4-carboxy aldehyde (4' -aldehyde-4-carboxyl dibenzene, referred to as AC-DBZ, structure _{CHO (C 6 H 6) 2} COOH) aqueous solution (100 mM After the shaking was uniformly reacted for 2 hours, the substrate 105 as shown in the reaction shown in Fig. 3C was obtained. The aldehyde group (-CHO) at one end of AC-DBZ can be coupled with the primary amine group (-NH ₂ ) on the substrate, and the carboxyl group (-COOH) at the other end can be reserved as an affinity agent, which is reserved for the reaction. Derivation of biomolecular probes. Thereafter, the wafers were each washed with dichloromethane and ionized water for 10 minutes to remove excess AC-DBZ molecules on the wafer surface.

It is noted that the substrate 103 in Figure 3A is a semi-finished wafer of antibody probe wafers. 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 of the anchor molecules on the substrates 104 and 105, and the antibody probes required for any sensing use have not yet been coupled. Such semi-finished substrates 104 and 105 can be stored under suitable conditions for a long period of time. Typical for at least three months. When desired, the semi-finished substrates 104 and 105 can be further processed rapidly, linking any of a variety of antibody probes suitable for covalent bonding to a carboxyl group for use in a particular detection application.

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. In medical applications, 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 taken out of stock. Quickly perform probe coupling processing to quickly and massively produce electrical detection antibody wafers for real-time tracking of disease viruses.

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 target probes for detecting specific antigens for related biomedicine. 111 (target) detection in samples (blood, urine, body fluids, 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 conductivity molecule thiophene sulfide: 5'-(methyl)thiol group- 5-alcohol-2,2'-dithiophene (5"-mercaptomethyl-5-aldehyde-2,2'-dithiophene (MMAD). The aldehyde group [-CHO] of this organic conductivity molecule can be combined with an antibody probe ( The amine group [-NH ₂ ] on the probe) is subjected to an imine bonding (shiff base) reaction to covalently bond the antibody probe to the organic conductive substrate 101. Thereafter, the target substance ( Target) The ability to integrate the ability to measure the wafer substrate 111.

a. HBS buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO) ₄ , 2 mM KH ₂ PO ₄ ) A constant flow rate (5 μl/min) was introduced into the flow path, and passed through the wafer substrate 101 containing the 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, injected into the flow channel, flowing through the wafer substrate 101, and subjected to imine bond bonding with the aldehyde group [-CHO] ( Imine bonding, shiff base) reaction, forming a covalent bond (-CH=N-). Covalently bonding an 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 in which the terminal probe was covalently bonded with the antibody probe-derived compound shown in Fig. 4 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 M) Immunoglobulin A), Immunoglobulin E, and Immunoglobulin D have the ability to specifically bind to the target, which facilitates biomedical testing related to electrical sensing in the future.

Example 2:

The chemical reaction of Figure 5 shows that the basic organic conductivity substrate 102, which can be adapted to link various antibody probes, can be linked to various antibody probes required for specific target detection for correlation. Detection of target in biomedical samples (blood, urine, body fluids, semen, saliva, laryngeal samples).

The chemical reaction of Fig. 5 shows that, in an embodiment of the invention, the surface of the base wafer substrate 102 of Fig. 2 is fixed with an organic conductive molecular oligobenzene sulfide: p -4'-thiol-4- Aldehyde-1,1-diphenyl (p-4 ' -mercapto-4-aldehyde dibenzene, abbreviated as MA-DBZ, structure CHO(C ₆ H ₆ ) ₂ COOH). The aldehyde group [-CHO] of the organic conductivity molecule can be reacted with an imine bonding (shiff base) on an amine [-NH ₂ ] on an antibody probe to covalently bind the antibody probe Bonded to the organic conductivity substrate 102. Thereafter, an inductive wafer substrate 112 having the ability to specifically bind to a target is formed.

a. HBS buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO) ₄ , 2 mM KH ₂ PO ₄ ) A constant flow rate (5 μl/min) enters the flow path and flows 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, injected into the flow channel, flowing through the wafer substrate 102, and subjected to imine bond bonding with the aldehyde group [-CHO] (imine Bonding, shiff base) reaction, forming a covalent bond (-CH=N-). Covalently bonding an antibody probe to the organic conductivity substrate 102

c. After the reaction is completed, the balance substrate 112 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 112 to which the terminal probe is covalently bonded with the antibody probe-derived compound shown in Fig. 5 can be 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 M) Immunoglobulin A), Immunoglobulin E, and Immunoglobulin D have the ability to specifically bind to the target, which facilitates biomedical testing 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 linked to various antibody probes required for specific target detection for correlation. Detection of target in biomedical samples (blood, urine, body fluids, semen, saliva, laryngeal samples).

The chemical reaction of Figure 6 shows that, in one embodiment of the invention, the surface of the base wafer substrate 104 of Figure 3B has an organic conductance oligothiophene sulfide: 5"-aldehyde-5-carboxy-2,2 ',5',2"-trithiophene (5"-aldehyde-5-carboxyl-2,2',5',2"-trithiophene, abbreviated as AC-TTP, the structure is CHO(C ₄ H ₂ S) ₂ COOH The organic conductivity molecule formed by the use of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (N-ethyl-N'-(dimethylaminopropyl)carbodiimide in the chemical reaction of Figure 6 Abbreviated as E-DAPC), with N-hydroxysuccinimide (HIS), [HO-N-(CO) ₂ (CH ₂ ) ₂ )] for the anchoring of carboxylic acid groups of organic conducting molecules [ -COOH] An amidation reaction is carried out 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 schemes of Figure 6, respectively. This 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 or the like:

a. HBS buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO) ₄ , 2 mM KH ₂ PO 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. 50 microliters of a 0.1 M HIS/0.4 M 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 microliters of a solution of antibody molecules having an amine group concentration of about 200 micrograms/ml, injected into the flow channel, flowing through the wafer substrate 104 to activate the group with the carboxyl group (-CO-OC ₄ O ₂ H ₄ N The amidation reaction is carried out to form a covalent bond (-CONH-). An antibody probe is covalently bonded to the organic conductivity substrate 104.

d. After the reaction is completed, the substrate is washed with a constant flow of HBS buffer, and then 50 μl of a solution of ethanolamine hydrochloride pH 8.5 having a concentration of about 1 M is injected into the flow channel and flowed through the wafer substrate 114 so as to be unreacted. The activating group (-CO-OC ₄ O ₂ H ₄ N) is completely amidated. At this time, the substrate 114 in which the terminal molecule was covalently bonded with the antibody molecule-derived compound shown in Fig. 6 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 M) Immunoglobulin A), Immunoglobulin E, and Immunoglobulin D have the ability to specifically bind to the target, which facilitates 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 linked to various antibody probes required for specific target detection for correlation. Detection of target in biomedical samples (blood, urine, body fluids, semen, saliva, laryngeal samples). FIG chemical reaction of 7 is displayed, in a preferred embodiment of the present invention among the basic bio-surface of the substrate wafer 105 of FIG. 3C, oligonucleotides having benzene (Phenyl) sulfide: 4 '- biphenyl-4-carboxy aldehyde (4 '-aldehyde-4-carboxyl dibenzene, referred to as AC-DBZ, structure _{CHO (C 6 H 6) 2} COOH) composed of electrically conductive organic molecules in a chemical reaction in the FIG 7, using N- succinimidyl-yl diameter N-ethyl-N'-(dimethylaminopropyl)carbodiimide, abbreviated as E-DAPC, and N-hydroxysuccinimide, referred to as HSI, [HO-N-(CO) ₂ (CH ₂ ) ₂ ) The amidation reaction of the carboxylic acid group [-COOH] of the anchoring organic conductivity molecule AC-DBZ is carried out to covalently bond the antibody probe to the organic conductivity substrate 105. The molecular structures of HSI and E-DAPC are shown in the reaction schemes of Figure 7, respectively. 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 or the like:

a. HBS buffer (10 mM HEPES pH 7.4, 150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO) ₄ , 2 mM KH ₂ PO ₄ ) 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. 50 microliters of a 0.1 M HIS/0.4 M E-DAPC (1:1) mixed solution was injected into the flow path and passed through the wafer substrate 105 to activate the carboxylic acid group of AC-DBZ.

c. 50 μl of a biomolecule solution having an amine group concentration of about 200 μg/ml is injected into the flow channel and passed through the wafer substrate 105 to activate the carboxyl group with the carboxyl group (-CO-OC ₄ O ₂ H ₄ N The amidation reaction is carried out 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, and then 50 μl of a solution of ethanolamine hydrochloride pH 8.5 having a concentration of about 1 M is injected into the flow channel and flowed through the wafer substrate 115 so as to be unreacted. The activating group (-CO-OC ₄ O ₂ H ₄ N) is completely amidated. At this time, the substrate 115 to which the terminal probe is covalently bonded with the antibody probe-derived compound shown in Fig. 7 can be 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 M) Immunoglobulin A), Immunoglobulin E, and Immunoglobulin D have the ability to specifically bind to the target, which facilitates 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 as an example, the present invention has an antibody probe wafer with a covalently bonded organic conductivity anchoring compound, and as can be understood, it can also be applied to other uses. Gold, silver, copper, platinum, nickel, zinc, bismuth, mercury or palladium are treated as different treatments for 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, it is to be understood that the scope of the present invention is defined by the scope of the appended claims.

Example 5:

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 performing a related biometric medical sample (blood, Affinity electrical detection in urine, body fluids, semen, saliva, and laryngeal samples.

The chemical reaction of Fig. 8 shows that, in one embodiment of the present invention, the basic immunoglobulin G antibody molecule 200 of Fig. 8 has a surface covalently bonded organic having the ability to conduct electrons. An organic electron-conducting oligomer 301, for example, a preferred embodiment thereof is 3-(thiophen-2-yl)propanal [(C ₄ H ₃ S) )-(CH ₂ )-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 immunoglobulin E (Immunoglobulin E), the amine group [-NH ₂ ] on the 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 inductive antibody probe 501 having the ability to specifically bind to a target is formed.

a. HBS (10 mM HEPES pH 7.4) buffer (150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO ₄ , 2 mM KH ₂ PO ₄ ) A constant flow rate (5 μl/min) enters the flow channel and flows through and cleans the wafer 200 to which the antibody probe is immobilized.

b. 50 μl of a 3-(thiophen-2-yl)propanal organic conductance oligo molecule solution 301 having an aldehyde group of about 10 mM, injected into the flow channel, and slowly flowing through the antibody probe wafer 200 for 24 hours. The imine bond reaction is carried out with four amine groups on the surface of the antibody molecule, and 3-(thiophen-2-yl)propanal is covalently bonded to the antibody probe molecule.

c. After the reaction is completed, the equilibrium antibody probe wafer substrate 501 is washed with a constant flow of HBS or PBS buffer to flow the finally reacted 3-(thiophen-2-yl)propanal oligo molecule 301 out of the wafer substrate. At this time, the antibody probe wafer substrate 501 having the surface covered with the 3-(thiophen-2-yl)propanal organic conductive oligo molecule 301 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 electrical sensing in the future.

Example 6:

The chemical reaction of Figure 9 shows one of the embodiments of the invention required to prepare a conductive electrons antibody probe 502 for performing related inductive biomedical samples (blood, urine, body fluids). , sperm, saliva, laryngeal fluid) in the target (affinity) electrical detection.

The chemical reaction of Figure 9 shows that, in one embodiment of the invention, the basic immunoglobulin G antibody molecule 200 of Figure 9 is covalently bonded to the surface to coat the organic conductance of the conducting electrons. An organic electron-conducting oligomer 302, for example, a preferred embodiment thereof is 3-phenylpropanal [(C ₆ H ₅ )-(CH ₂ )-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 immunoglobulin E (Immunoglobulin E), the amine group [-NH ₂ ] on the immunoglobulin D (Immunoglobulin D) undergoes an imine bonding (shiff-base reaction) reaction to the organic conducting oligo molecule 302 Covalently bonded to antibody probe 200. Thereafter, an inductive antibody probe 502 having the ability to specifically bind to a target is formed.

a. HBS (10 mM HEPES pH 7.4) buffer (150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO ₄ , 2 mM KH ₂ PO ₄ ) A constant flow rate (5 μl/min) enters the flow channel and flows through and cleans the wafer 200 to which the antibody probe is immobilized.

b. 50 microliters of a 3-phenylpropanal organic conductance oligo molecule solution 302 having an aldehyde group of about 10 mM, injected into the flow channel, and slowly flowing 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 equilibration antibody probe wafer substrate 502 is washed with a constant flow of HBS or PBS buffer to flow unreacted 3-phenylpropanal oligo molecule 302 out of the wafer substrate. At this time, the antibody probe wafer substrate 502 having the surface covered with the 3-phenylpropanal organic conductive oligo molecule 302 as shown in Fig. 9 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 electrical sensing in the future.

Example 7:

The chemical reaction of Figure 10 shows that one of the embodiments of the present invention required to prepare a conductive electrons antibody probe 503 is used to perform biosensory samples (blood, urine, body fluids). , sperm, saliva, laryngeal fluid) in the target (affinity) electrical detection.

The chemical reaction of Figure 10 shows that, in one embodiment of the invention, the basic immunoglobulin G antibody molecule 200 of Figure 10, covalently bonded to the surface, has organic conductance with conducting electrons. An organic electron-conducting oligomer 303, for example, a preferred embodiment thereof is 3-(1-pyrrol-2-yl)propanal (3-(1-pyrrol-2-yl)propanal[2-(C ₄ H ₄ N)-(CH ₂ )-CHO]. The aldehyde group [-CHO] of this organic conductivity oligo molecule can be combined with an antibody probe such as immunoglobulin G (immunoglobulin G), immunoglobulin M (Immunoglobulin M), immunoglobulin A (Immunoglobulin A), immunoglobulin E (Immunoglobulin E), immunoglobulin D (Immunoglobulin D) on the amine group [-NH ₂ ] for imine bonding (imine bonding, The shiff-base reaction is carried out to covalently bond the organic conducting oligo molecule 303 to the antibody probe 200. Thereafter, an inductive antibody probe 503 having a specific binding ability to a target is formed. .

a. HBS (10 mM HEPES pH 7.4) buffer (150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO ₄ , 2 mM KH ₂ PO ₄ ) A constant flow rate (5 μl/min) enters the flow channel and flows through and cleans the wafer 200 to which the antibody probe is immobilized.

b. 50 μl of a 3-(1-pyrrol-2-yl)propanal organic conducting oligo molecule solution 303 having an aldehyde group of about 10 mM, injected into the flow channel, and slowly flowing through the antibody probe wafer 200, continuing After 24 hours, it was subjected to an imine bond-bonding reaction with four amine groups on the surface of the antibody molecule, 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 finally reacted 3-(1-pyrrol-2-yl)propanal oligo molecule 303 out of the wafer substrate. At this time, the antibody probe wafer substrate 503 having the surface covered with the 3-(1-pyrrol-2-yl)propanal organic conductive oligo molecule 303 shown in Fig. 10 was obtained.

This type of conductance antibody sensing wafer has the ability to specifically bind to an antigen target, facilitating biomedical testing associated with electrical sensing in the future.

Example 8:

The chemical reaction of Figure 11 shows one of the embodiments of the invention required to prepare a conductive electrons antibody probe 504 for performing related inductive biomedical samples (blood, urine, body fluids). , sperm, saliva, laryngeal fluid) in the target (affinity) electrical detection.

The chemical reaction of Figure 11 shows that, in one embodiment of the invention, the basic immunoglobulin G antibody molecule 200 of Figure 11 is covalently bonded to the surface to coat the organic conductance of the conducting electrons. An organic electron-conducting oligomer 304, for example, a preferred embodiment thereof is 3-(pyridin-2-yl)propanal (2-(pyridin-2-yl) propanal) [2-(C ₅ H ₄ N)-(CH ₂ )-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, Immunoglobulin E (Immunoglobulin E), an amine group [-NH ₂ ] on immunoglobulin D (Immunoglobulin D) undergoes an imine bonding (shiff-base reaction) reaction to expose the organic conducting oligo molecule 304 Covalently bonded to antibody probe 200. Thereafter, an inductive antibody probe 504 having the ability to specifically bind to a target is formed.

a. HBS (10 mM HEPES pH 7.4) buffer (150 mM NaCl, 3.4 mM EDTA, 0.005% surfactant P20) or PBS (Phosphate buffered saline pH 7.4) buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na ₂ HPO ₄ , 2 mM KH ₂ PO ₄ ) A constant flow rate (5 μl/min) enters the flow channel and flows through and cleans the wafer 200 to which the antibody probe is immobilized.

b. 50 microliters of a 3-(pyrrole-2-yl)propanal organic conductivity oligo molecule solution 304 having an aldehyde group of about 10 mM, injected into the flow channel, and slowly flowing through the antibody probe wafer 200 for 24 hours. In an hour, it is subjected to an imine bond reaction with four amine groups on the surface of the antibody molecule, and 3-(pyrrole-2-yl)propanal is covalently bonded to the antibody probe molecule.

c. After the reaction is completed, the equilibration antibody probe wafer substrate 504 is washed with a constant flow of HBS or PBS buffer so that the finally reacted 3-(pyrrole-2-yl)propanal oligo molecule 304 flows out of the wafer substrate. At this time, an antibody probe wafer substrate 504 having a surface covered with 3-(pyrrole-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 electrical sensing in the future.

Although the present invention has been described above with reference to the drawings in the preferred embodiments, the present invention is not intended to limit the invention. For example, although the description of the present invention is described in detail by taking an electrically conductive antibody probe wafer as an example, the surface of the present invention is coated with an organic conductive oligo molecule (1) 3-(thiophen-2-yl)propanal, (2)3- An antibody probe for phenylpropanal, (3) 3-(1-pyrrol-2-yl)propanal, (4) 3-(pyrrole-2-yl)propanal, as can be understood, as shown in Figure 12 Also shown in -15, it is also applicable to derivatives of the other four organic conducting oligo molecules described above to prepare antibody probes having conducting electrons.

Further, although the present invention has been described above with reference to the drawings in the preferred embodiments, the present invention is not intended to limit the invention. As can be appreciated, as shown in Figures 16-21, the same can be applied to other organic conducting oligo molecules and derivatives thereof having conducting electrons to prepare antibody probes with conducting electrons. Applications include, for example, electrical sensing wafers, electrochemical sensing wafers, field effect transistor sensing wafers, piezoelectric sensing wafers, and the like. Therefore, it is to be understood that the scope of the present invention is defined by the scope of the appended claims.

110‧‧‧ wafer substrate

120‧‧‧ wafer gold film electrode

100‧‧‧ Gold Film Wafer

103‧‧‧Gold film wafer with joints

101,102,104,105‧‧‧Organic Conductance Anchoring Molecules

111,112,114,115‧‧‧ Conductance Antibody Probe Wafer

200‧‧‧Basic antibodies

301,302‧‧‧Organic Conducting Molecules (Conductive Leciting Molecules)

501,502,503,504‧‧‧ Conducting antibodies

The chemical reactions of Figures 1, 2 and 3 respectively show that a ring of a specific length is organically treated on the surface of a substrate coated with gold or silver by chemical modification according to a preferred embodiment of the present invention. The electrically conductive oligothiophene and the oligobenzene molecule are immobilized for attachment to a basic antibody probe wafer substrate of an antibody to be used.

The chemical reaction of Fig. 1 shows that on the wafer substrate coated with the gold (Au) metal thin film layer, the chemical modification system uses a covalent bond of sp3 (gold and sulfur atom) mixed orbital to form the organic conductivity molecular oligomer. An 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 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 p -4'-thiol-4-aldehyde-1,1-diphenyl. , fixed to the metal film.

The chemical reaction of Fig. 3A shows that an alkyl sulfide: 2-thiol-ethanolamine is immobilized on a wafer substrate coated with a vapor deposited gold or silver metal thin film layer.

The wafer chemical reaction of Figure 3B shows that on the surface of the antibody probe wafer substrate of Figure 3A, the organic conductive molecule oligothiophene sulfide derivative: 5"-aldehyde-5-carboxy-2,2',5', 2"-trithiophene (5"-aldehyde-5-carboxyl-2,2',5',2"-trithiophene), the aldehyde group (-CHO) at one end can be combined with the primary amine group on the substrate (-NH ₂ The coupling reaction is carried out, and the carboxyl group (-COOH) at the other end can be reserved as a affinity agent, and the derivative of the antibody molecule probe is retained for the reaction.

FIG. 3C show a chemical reaction of a wafer, the wafer substrate surface antibody probe of FIG. 3A, the use of an oligonucleotide molecule organic conductance phenyl sulfide derivative: 4 '- biphenyl-4-carboxy aldehyde (4' -aldehyde-4- Carboxyl dibenzene), the aldehyde group (-CHO) at one end can be coupled with the primary amine group (-NH ₂ ) on the substrate, and the carboxyl group (-COOH) at the other end can be reserved as a hydrophilic agent. Derivatization of antibody probes is performed.

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 subjected to an imine bonding reaction with an amine group [-NH ₂ ] on an antibody probe to covalently bond the antibody probe to On the organic 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 [-NH ₂ ] on an antibody probe (Ab probe) to covalently bond the antibody probe. Onto 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 [-NH ₂ ] on an antibody probe to covalently bond the antibody probe (probe). Bonded to the organic conductivity 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 conducting molecule can be subjected to an amidation reaction with an amine group [-NH ₂ ] on a basic antibody probe to share a basic antibody probe. The bond is 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 substrate through an organic electron-conducting moleecule having a conducting electrons 3-(thiophen-2-yl) Propionaldehyde and its derivatives (3-(thiophen-2-yl) propanal and derivatives). The aldehyde group [-CHO] of the organic conductivity molecule can be covalently bonded to the amine group [-NH ₂ ] on the antibody probe for imine bonding to coat the organic conductivity molecule to the antibody probe. on.

The chemical reaction of Figure 9 shows that the conductivity antibody probe flows through the surface of the basic antibody probe wafer substrate through the 3-phenylpropanal and derivatives of the organic conductivity molecule having conductive electrons. The aldehyde group [-CHO] of the organic conductivity molecule can be covalently bonded to the amine group [-NH ₂ ] on the antibody probe for imine bonding, so that the organic conductivity molecule can be coated onto the basic antibody probe. On the needle.

The chemical reaction of Figure 10 shows the conductivity antibody probe flowing through the surface of the basic antibody probe wafer substrate through the organic conductive molecule 3-(1-pyrrol-2-yl)propanal and its derivatives (3- (1-pyrrol-2-yl)propanal and derivatives). The aldehyde group [-CHO] of the organic conductivity molecule can be covalently bonded to the amine group [-NH ₂ ] on the antibody probe for imine bonding to coat the organic conductivity molecule to the antibody probe. on.

The chemical reaction of Figure 11 shows that 3-(pyrrole-2-yl)propanal and its derivatives (3-(pyridin-2-) are passed through the surface of the basic antibody probe wafer substrate through the organic conducting molecule possessing conductive electrons. Yl) propanal and derivatives). The aldehyde group [-CHO] of the organic conductivity molecule can be covalently bonded to the amine group [-NH ₂ ] on the antibody probe for imine bonding to coat the organic conductivity molecule to the antibody probe. on.

Figure 12 is an organic conductivity molecule 3-(thiophen-2-yl)propanal and its derivative 3-(thiophen-2-yl)propanal and derivatives, which are applicable to the above-mentioned conductivity probe application of the present invention, to have Conducting electron probe antibody for conducting electrons.

Figure 13 is a 3-phenylpropanal and derivatives of the organic conductivity molecule 3-phenylpropanal and its derivatives which are suitable for use in the above-described electroconductive antibody probe of the present invention to prepare a conducting antibody probe having electron conduction.

Figure 14 is an organic conductivity molecule 3-(1-pyrrol-2-yl)propanal and its derivative 3-(1-pyrrol-2-yl)propanal and derivatives which are applicable to the use of the above-described electroconductive antibody probe of the present invention. To prepare a conducting antibody probe with conducting electrons.

Figure 15 is an organic conductivity molecule 3-(pyrrole-2-yl)propanal and its derivative 3-(pyridin-2-yl)propanal and derivatives which can be suitably used for the above-mentioned electroconductive antibody probe of the present invention to prepare Conductive antibody probe with electron conduction.

Figure 16 is an organic conductivity molecule cis-4-phenyl-3-butenal and its derivative ( E )-4-phenylbut-3-enal and derivatives which are suitable for use in the above-described conductivity probe of the present invention. To prepare a conducting antibody probe with conducting electrons.

Figure 17 is an organic conductivity molecule cis-4-(pyridin-2-yl)-3-butenal and its derivative ( E )-4-(pyridin-) which can be suitably used for the above-mentioned conductivity probe of the present invention. 2-yl)but-3-enal and derivatives to prepare a conducting antibody probe having electron conduction.

Figure 18 is an organic conductivity molecule cis-hepta-4,6-dienal and its derivative ( E )-hepta-4,6-dienal and derivatives which are applicable to the above-mentioned conductivity probe application of the present invention. A conductivity probe probe having electron conduction is prepared.

Figure 19 is an organic conductivity molecule trans-seven-4,6-dienal and its derivatives ( Z )-hepta-4,6-dienal and derivatives which are applicable to the above-mentioned conductivity probe of the present invention. A conductivity probe probe having electron conduction is prepared.

Figure 20 is an organic conductivity molecule cis-4-(cyclohex-2-enyl)butanal and its derivative ( E )-4-(cyclohex-2-) which can be suitably used for the above-mentioned electroconductive antibody probe of the present invention. Enylidene) butanal and derivatives to prepare a conducting antibody probe with conducting electrons.

Figure 21 is an organic conductivity molecule 2-(cyclohydro-2-yl)acetaldehyde and its derivative 2-(naphthalen-2-yl)acetaldehyde and derivatives which are applicable to the above-mentioned conductivity probe application of the present invention. Conductive antibody probe with electron conduction.

It should be noted that, for the novelty of the present invention, the invention has been described in detail above with reference to the preferred embodiments illustrated in the drawings. In the sheet system, one of several possible ways of performing electron conduction, that is, the organic conductive molecule is also used as a linker of the antibody counter electrode, that is, as depicted in Figures 4, 5, 6 and 7. However, as will be appreciated by those skilled in the art, other arrangements of organic conductivity molecules (conductive attracting molecules) in the system of the present invention are equally possible and feasible. For example, a slightly different arrangement encompasses 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.

110. . . Wafer substrate

120. . . Wafer gold film electrode

101,102. . . Organic conductance anchor molecule

111,112. . . Conductive antibody probe chip

104,105. . . Organic conductance anchor molecule

114,115. . . Conductive antibody probe chip

200. . . Basic antibody

301,302. . . Organic conducting molecule (conductive attracting molecule)

501,502. . . Conductive antibody

Claims (18)

A conductive antibody probe wafer comprising: a substrate having a metal film layer deposited on a surface thereof; a plurality of organic conductance anchor molecules, wherein the organic conductance anchor molecules are cyclic chain molecules, each Each of the organic conductance anchor molecules is directly covalently bonded to the metal thin film layer by one end of one of the cyclic chain molecular structures, or indirectly covalently bonded to the metal thin film layer through the linker; And a plurality of basic antibody probes, each of the basic antibody probes covalently bonded to the organic conductive derivative anchor molecules corresponding to a certain anchor molecule against the first end of the cyclic chain molecular structure a two-terminal; and a plurality of conductivity probes, wherein the plurality of organic conductivity molecules, substantially having an extended and substantially circular chain, are covalently bonded to the base antibody probe, each of the conductance Each of the antibody probes is covalently bonded to the second end of the first end of the organic chain of the organic conductive derivative anchor molecules corresponding to the anchor chain molecule to form a straight line from the metal film layer To the antibody Linear vertical conductive electrode a whole antigen-binding site (antigen-binding site), wherein such organic molecules as the organic electrically anchoring oligothiophene (oligo-thiophene) sulfide structure [HS- (CH _{2) n} - (C ₄ H ₂ S)-(C ₄ H ₂ S) _m -X], wherein n=0~12 does not include 8~12, m=0~3, X series is aldehyde group [-CHO], carboxylic acid a group [-COOH], a disulfide bond [-SS-], an amine group [-NH ₂ ] or a thiol group [-SH], an organic oligo-benzene sulfide having a structure of [HS-(CH _{2 ) n} -(C ₆ H ₄ )-(C ₆ H ₄ ) _m -X], wherein n=0~12 does not include 8~12, m=0~3, X series is aldehyde group [-CHO], carboxy Acid group [-COOH], disulfide bond [-SS-], amine group [-NH ₂ ] or thiol group [-SH], organic oligothiophene sulfide derivative, structure [CHO-(CH ₂ ) _n -(C ₄ H ₂ S)-(C ₄ H ₂ S) _m -X], wherein n=0~12 does not include 8~12, m=0~3, X series is aldehyde group [-CHO], carboxy Acid group [-COOH], disulfide bond [-SS-], amine group [-NH ₂ ] or thiol group [-SH], and organic oligobenzene sulfide derivative, structure [CHO-(CH ₂ ) _n -(C ₆ H ₄ )-(C ₆ H ₄ ) _m -X], wherein n=0~12 does not include 8~12, m=0~3, X series is aldehyde group [-CHO], carboxylic acid Base [-COOH], disulfide bond [-SS-], amine group [-NH ₂ ] or thiol group [-SH], one of them. The electroconductive antibody probe wafer of claim 1, wherein the organic electrically conductive anchor molecule which is an organo oligothiophene sulfide is directly covalently bonded to the metal thin film layer at the first end. The electroconductive antibody probe wafer of claim 1, wherein the organic conductance anchor molecule which is an organo oligothiophene sulfide is directly covalently bonded to the antibody probe at its second end. For example, the conductivity antibody probe chip of claim 1 of the patent scope, wherein The organic electrically conductive anchor molecule of the oligobenzene sulfide can be directly covalently bonded to the metal thin film layer at the first end. The electroconductive antibody probe wafer of claim 1, wherein the organic conductance anchor molecule which is an organic oligobenzene sulfide is directly covalently bonded to the antibody probe at the second end thereof. The electroconductive antibody probe wafer of claim 1, wherein the organic electrically conductive anchor molecule is covalently bonded to the metal thin film layer through a coupling, wherein the organic electrically conductive anchor molecule is an organic oligothiophene sulfide a derivative or an organic phenyl sulfide derivative, the linkage being an organoalkyl sulfide having the structure [HS-(CH ₂ ) _n -NH ₂ ], wherein n=1 to 7, and the organic conductance anchor The total bonded carbon number of the molecule and the linker does not include more than 8 carbon chains. The electroconductive antibody probe chip of claim 6, wherein the organic conductance anchor molecule of the organo oligothiophene sulfide derivative is indirectly covalently bonded to the first end of the organic conductance anchor molecule through the linker Attached to the metal film layer. The electroconductive antibody probe wafer according to claim 6, wherein the organic conductance anchor molecule which is an organic oligothiophene sulfide derivative may be covalently bonded to the basic antibody probe at the second end thereof. The electroconductive antibody probe chip according to claim 6 , wherein the organic electrically conductive anchor molecule which is an organic oligobenzene sulfide derivative is indirectly covalently bonded to the first end of the organic conductance anchor molecule through the linker Attached to the metal film layer. For example, the conductivity antibody probe chip of claim 6 of the patent scope, wherein The organic conductance anchor molecule of the oligobenzene sulfide derivative can be covalently bonded to the base antibody probe at its second end. The electroconductive antibody probe wafer of claim 1, wherein a plurality of organic conducting molecules are covalently bonded to the basic antibody probe, wherein the organic conducting molecule is organic 3-(thiophene-2). -yl)propanal and its derivative 3-(thiophen-2-yl)propanal and derivatives, the structure is [H-(C ₄ H ₂ S) _n -(CH ₂ ) _m -X], where n=1, 2, 3, 4, m = 0 to 12 does not include 4 to 12. The conductive antibody probe wafer of claim 11, wherein the organic conductive molecule having the structure [H-(C ₄ H ₂ S) _n -(CH ₂ ) _m -X] may have one end functional group -X Covalently bonded to a basic antibody probe, the functional group -X is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [-NH ₂ ] or sulfur Alcohol group [-SH]. The electroconductive antibody probe wafer of claim 1, wherein a plurality of organic conducting molecules are covalently bonded to the basic antibody probe, wherein the organic conducting molecule is organic 3-phenylpropanal. And its derivatives (3-phenylpropanal and derivatives), the structure is [H-(C ₆ H ₄ ) _n -(CH ₂ ) _m -X], where n=1, 2,3,4, m=0~12 Does not include 4~12. The electroconductive antibody probe wafer of claim 13, wherein the organic conducting molecule having the structure [H-(C ₆ H ₄ ) _n -(CH ₂ ) _m -X] may have one end functional group -X and The basic antibody probe is covalently bonded, and the functional group -X is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [-NH ₂ ] or a thiol. Base [-SH]. For example, in the radioactive antibody probe chip of claim 1, wherein a plurality of organic conducting molecules are covalently bonded to the basic antibody probe, wherein the organic conducting molecular system is (1) 4-phenyl-3-butenal and its derivatives ( E )-4-phenylbut-3-enal and derivatives[H-(C ₆ H ₄ -C ₂ H ₂ ) _n -(CH ₂ ) _m -X], where n=1, 2, 3, 4, m=0~12 does not include 4~12. The electroconductive antibody probe wafer of claim 15 wherein the organic conducting molecule having the structure [H-(C ₆ H ₄ -C ₂ H ₂ ) _n -(CH ₂ ) _m -X] is respectively The one-end functional group -X is covalently bonded to an antibody probe, which is an aldehyde group [-CHO], a carboxylic acid group [-COOH], a disulfide bond [-SS-], an amine group [-NH ₂ ] or thiol group [-SH]. The electroconductive antibody probe wafer according to any one of claims 1 to 10, wherein the metal thin film layer is a copper, platinum, zinc, ruthenium, mercury or palladium thin film layer. The electroconductive antibody probe wafer according to any one of claims 1 to 16, wherein the metal thin film layer is a microarray metal thin film layer.