CN210894379U - Antibody detection system based on micro-fluidic chip - Google Patents

Abstract

The utility model provides an antibody detecting system based on micro-fluidic chip, including affinity chromatography post and micro-fluidic chip, affinity chromatography post is protein G affinity chromatography post or protein A affinity chromatography post, micro-fluidic chip includes piece and lower piece, the piece closely the pressfitting in the upper surface of lower piece, the piece of going up is seted up and is link up the application of sample hole of piece, the bottom of affinity chromatography post is inserted in the application of sample hole. The utility model discloses make up high flux, high sensitivity's micro-fluidic chip and the efficient affinity chromatographic column of purification, utilize the affinity chromatographic column to carry out one step of purification with the antibody in the blood sample, carry out the further detection of micro-fluidic chip to the antibody of purification again, can not only improve the sensitivity and the specificity that the antibody detected, solved the problem that the blood sample blockked up the capillary microchannel of micro-fluidic chip simultaneously.

Description

Antibody detection system based on micro-fluidic chip

Technical Field

The utility model belongs to the technical field of it is immune, especially, relate to an antibody detecting system based on micro-fluidic chip.

Background

The micro-fluidic chip technology is a technology for controlling micro-scale liquid developed in the last two thirty years. The micro-fluidic chip technology integrates comprehensive technologies in the fields of engineering, physics, chemistry, micro-processing, biological engineering and the like. Meanwhile, as a new scientific technology, the microfluidic chip technology has received wide attention from researchers in various fields, such as physical science, life science, and engineering science.

The demands of scientific research markets and medical treatment and the increasing maturity of micro-nano processing technology promote the generation of the micro-fluidic chip technology and accelerate the development thereof, so that the micro-fluidic chip technology also occupies an important position in the field of clinical medicine, especially in the aspect of clinical medicine inspection. At present, the microfluidic chip technology is widely applied to the detection of bacteria and viruses, and is also applied to immune markers, genetic diseases and tumor markers of autoimmune diseases to play a role in single detection or combined detection. With the introduction of technologies such as microelectronics, materials science, bioscience, pharmacy, and clinical medicine, biochip technology will be in clinical medicine examination or will be indispensable in the near future.

Compared with the traditional biotechnology and disease diagnosis, the micro-fluidic chip has the main characteristics that: the liquid flow is controllable, the consumption of samples and reagents is very little, the analysis speed is high, the pollution is small, hundreds of samples can be analyzed in a short time (within minutes or even shorter time), and the whole processes of pretreatment and analysis of the samples can be realized on line. Meanwhile, the microfluidic chip is easy to array, so that high-throughput, system integration, miniaturization, automation and portable detection can be realized. The microfluidic chip comprises a non-flow static miniature experimental system, and analysis is generally carried out by detecting different reactions on a dot matrix; and a micro experimental system by means of an external force such as an electromagnetic force, a centrifugal force, and the like, and analysis of a sample is generally achieved by magnetic particles, a centrifugal device, and the like.

The rapid development of microfluidic chips requires not only a sensitive detection format to be adapted to it, but also a sample type to be matched with it for support. The detection area of the chip is very small, so the requirement on the sample is high, and in general, various anticoagulant plasma, various coagulant serum and whole blood can be used as microfluidic samples to be detected. However, because the components in blood are complex, during the storage process, impurities such as flocculent precipitates and the like are easily formed, so that the non-specific reaction occurs in the experimental result or the detection cannot be normally carried out due to the blockage of a channel.

Protein G is a cell surface protein from the streptococcal G family, a type three Fc receptor, which binds to the Fc portion of antibodies through a non-immune mechanism similar to that of protein a. Under certain specific conditions, protein G is capable of binding all antibodies of the IgG subclass of human origin. Protein G is coupled to an agarose matrix as an affinity ligand, can be specifically combined with antibody molecules in a sample, and other hybrid proteins flow through due to lower serum protein combination level, so that the protein G has extremely high selectivity, and the purity of over 95 percent can be achieved by one-step affinity chromatography. At present, affinity filler using Protein G as ligand is widely used for separation and purification of antibody, and has become a conventional and necessary medium for antibody production.

The antibody detection plays an important role in clinical detection, and is widely applied to the aspects of autoimmune disease detection, drug detection, antibody detection of various pathogenic bacteria and autoantibody detection. At present, the main methods for detecting antibodies clinically comprise ELISA (enzyme-linked immunosorbent assay), immunofluorescence, colloidal gold, colloidal selenium, Western-blot and the like. However, the antibodies have high content of blood environmental proteins and are various, and the removal of proteins by an aggressive chemical method cannot be performed, so that the sensitivity and specificity of detection are affected. Therefore, the application of the microfluidic chip in the aspect of clinical antibody detection is limited.

SUMMERY OF THE UTILITY MODEL

In view of this, the utility model aims at providing an antibody detecting system based on micro-fluidic chip, this system makes up high flux, high sensitivity's micro-fluidic chip and the efficient affinity chromatographic column of purification, utilizes the affinity chromatographic column to carry out further purification with the antibody in the blood sample, carries out the further detection of micro-fluidic chip to the antibody of purification again, can not only improve the sensitivity and the specificity that the antibody detected, has solved the problem that the blood sample blockked up micro-fluidic chip's capillary microchannel simultaneously.

In order to achieve the above purpose, the technical scheme of the utility model is realized like this:

the utility model provides an antibody detecting system based on micro-fluidic chip, includes affinity chromatography column and micro-fluidic chip, affinity chromatography column is albumen G affinity chromatography column or albumen A affinity chromatography column, micro-fluidic chip includes upper segment and lower piece, the inseparable pressfitting of upper segment in the upper surface of lower piece, the sample application hole of lining up the upper segment is seted up to the upper segment, the bottom of affinity chromatography column is inserted in the sample application hole.

Furthermore, the affinity chromatography column comprises a column tube, the top end of the column tube is provided with a sample adding port, the bottom end of the column tube is provided with a liquid discharging nozzle, and the inside of the column tube is sequentially provided with a sand core filter sheet I, a filler and a sand core filter sheet II from top to bottom.

Furthermore, the lower surface of the upper plate is provided with an upper detection unit, the upper detection unit comprises a capillary micro-channel and a waste liquid tank, one end of the capillary micro-channel is connected with the sample adding hole, and the other end of the capillary micro-channel is connected with the waste liquid tank.

Further, the lower surface of the upper piece is provided with at least two upper detection units, the upper detection units are circumferentially distributed on the outer sides of the sample adding holes by taking the centers of the sample adding holes as circle centers, each upper detection unit comprises a capillary micro-channel and a waste liquid tank, one end of the capillary micro-channel is connected with the sample adding holes, and the other end of the capillary micro-channel is connected with the waste liquid tank.

Furthermore, a sample neutralization area is arranged at the position, right opposite to the sample adding hole, of the upper surface of the lower piece, a lower detection unit is arranged at the position, right opposite to the upper detection unit, of the upper surface of the lower piece, the lower detection unit comprises a fluorescence mark area, a reaction detection point area, a reaction quality control point area and a waste liquid area which are sequentially arranged along the sample flowing direction, the fluorescence mark area, the reaction detection point area and the reaction quality control point area are right opposite to the capillary micro-channel, and the waste liquid area is right opposite to the waste liquid groove.

Furthermore, a microstructure area is arranged on the outer side of the sample neutralization area, the microstructure area is circular and concentric with the sample neutralization area, and a plurality of protrusions which are columnar or pyramid-shaped are uniformly distributed at intervals in the microstructure area.

Furthermore, the top edge of the sampling hole vertically extends upwards to form a guide hollow column, and an aluminum foil is sealed at the top of the guide hollow column.

Further, the volumes of the capillary micro-channel and the waste liquid groove are both 5-300 muL.

Furthermore, the micro-fluidic chip is made of quartz, glass or polyester substances and is circular or polygonal in shape.

Compared with the prior art, the antibody detection system based on the micro-fluidic chip has the following advantages:

1. the affinity chromatographic column is matched with the microfluidic chip for use, antibodies in samples of whole blood, plasma and serum are highly purified, so that the antibody detection in clinic can be conveniently, rapidly, highly sensitively and accurately analyzed, and then the microfluidic chip is used for detecting the samples.

2. The required amount of the sample is small, and the detection only needs 10-30 mu l generally.

3. The detection time is short, the operation is simple, and the full-automatic detection can be completed within 10-30 min.

4. The full closed system is designed with different number of detection devices, so that multiple detections of a single sample can be performed simultaneously, and high throughput of detection is realized.

5. The detection is a homogeneous reaction system, and the detection result has high sensitivity and strong specificity.

Drawings

The accompanying drawings, which form a part hereof, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention without undue limitation. In the drawings:

fig. 1 is a schematic diagram of an overall structure of an antibody detection system based on a microfluidic chip according to an embodiment of the present invention;

fig. 2 is a schematic diagram of a micro-fluidic chip of an antibody detection system based on a micro-fluidic chip according to an embodiment of the present invention;

fig. 3 is a lower schematic view of a microfluidic chip of an antibody detection system based on the microfluidic chip according to an embodiment of the present invention;

fig. 4 is a top plan view of a microfluidic chip of an antibody detection system based on the microfluidic chip according to an embodiment of the present invention, the microfluidic chip having four upper detection units;

fig. 5 is a top view of a microfluidic chip of an antibody detection system based on the microfluidic chip according to an embodiment of the present invention, the microfluidic chip having six upper detection units.

Description of reference numerals:

1. an affinity chromatography column; 11. a column tube; 12. a sand core filter disc I; 13. a filler; 14. a second sand core filter disc; 15. a sample addition port; 16. a liquid discharge nozzle; 2. a microfluidic chip; 21. loading the wafer; 211. a sample application hole; 2111. Guiding the hollow column; 212. a capillary microchannel; 213. a waste liquid tank; 22. carrying out sheet discharging; 221. a sample neutralization zone; 222. a microstructure region; 2221. a protrusion; 223. a fluorescent labeling region; 224. a reaction detection point region; 225. a reaction quality control point region; 226. a waste liquid zone.

Detailed Description

It should be noted that, in the present invention, the embodiments and features of the embodiments may be combined with each other without conflict.

In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are used merely for convenience of description and for simplicity of description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, should not be construed as limiting the present invention. Furthermore, the terms "first", "second", etc. are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first," "second," etc. may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless otherwise specified.

The present invention will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.

An antibody detection system based on a microfluidic chip is shown in fig. 1 and comprises an affinity chromatography column 1 and a microfluidic chip 2. The affinity chromatographic column 1 is a protein G affinity chromatographic column or a protein A affinity chromatographic column. The microfluidic chip 2 comprises an upper plate 21 and a lower plate 22, the upper plate 21 is tightly pressed on the upper surface of the lower plate 22, a sample adding hole 211 penetrating the upper plate 21 is formed in the center of the upper plate 21, and the bottom end of the affinity chromatography column 1 is inserted into the sample adding hole 211.

The affinity chromatography column 1 comprises a column tube 11, wherein a sample adding port 15 is arranged at the top end of the column tube 11, and a liquid discharging nozzle 16 is arranged at the bottom end of the column tube 11. The column tube 11 is a hollow cylinder, a sand core filter disc I12, a filler 13 and a sand core filter disc II 14 are sequentially arranged inside the column tube 11 from top to bottom, the column tube 11, the sand core filter disc I12 and the sand core filter disc II 14 are purchased from Shenzhen comma biology Limited and have the model of solid-phase extraction hollow column SPE hollow column 1ml, and the filler 13 is a coupling product of protein G and curdlan. Wherein the protein G is a genetically modified protein G, the active site of the genetically modified protein G is the same as that of the protein G obtained by natural separation, the protein G is purchased from Jining Baishi microbial technology GmbH, the model is recombinant protein G SPG-03, the curdlan is agar gel sugar, the gel polysaccharide is purchased from Beijing Borxi technology GmbH, and the model is AG-Bercharose FF.

As shown in fig. 2, five upper detection units, four upper detection units (see fig. 4) and six upper detection units (see fig. 5) may be provided on the lower surface of the upper sheet 21. By designing the detection devices with different combinations, the joint detection with different combination modes can be realized. Go up the detecting element with the center of application of sample hole 211 is centre of a circle circumference distribution in the outside of application of sample hole 211, it includes capillary microchannel 212 and waste liquid groove 213 to go up the detecting element, capillary microchannel 212 one end with application of sample hole 211 is connected, the other end with waste liquid groove 213 is connected. The driving force for the sample flow of the microfluidic chip 2 is capillary suction.

As shown in fig. 3, a sample neutralization region 221 is provided on the upper surface of the lower sheet 22 at a position facing the well 211, and the sample neutralization region 221 is solidified with a phosphate buffer solution having a pH of 10. The upper surface of the lower sheet 22 is provided with a lower detection unit at a position facing the upper detection unit, the lower detection unit comprises a fluorescence mark area 223, a reaction detection point area 224, a reaction quality control point area 225 and a waste liquid area 226 which are sequentially arranged along the sample flowing direction, the fluorescence mark area 223, the reaction detection point area 224 and the reaction quality control point area 225 are all facing the capillary micro-channel 212, and the waste liquid area 226 is facing the waste liquid groove 213.

In order to promote the uniform mixing of the sample and the buffer system, as shown in fig. 3, a microstructure area 222 is disposed outside the sample neutralization area 221, the microstructure area 222 is annular and concentric with the sample neutralization area 221, and a plurality of columnar or pyramid-shaped protrusions 2221 are uniformly spaced in the microstructure area 222.

As shown in FIG. 1, the top edge of the sample application hole 211 extends vertically upward to form a hollow guiding column 2111, and the hollow guiding column 2111 is provided to facilitate the lower end of the drain nozzle 16 of the affinity chromatography column 1 to be inserted into the sample application hole 211 vertically and smoothly. The top of the guide hollow post 2111 is sealed with aluminum foil (not shown). When purifying the antibody in the sample, the affinity chromatography column 1 is not inserted into the microfluidic chip 2, and therefore, the sample addition hole 211 is sealed by the aluminum foil, and the sample addition hole 211 is prevented from being polluted. When detecting a sample, the affinity chromatography column 1 pierces the aluminum foil and is inserted into the sample adding hole 211 of the microfluidic chip 2 through the guide hollow column 2111.

The volumes of the capillary micro-channel 212 and the waste liquid tank 213 are both 5 to 300. mu.L.

The micro-fluidic chip 2 is made of quartz, glass or polyester substances and is circular or polygonal in shape.

The fluorescent labeling area 223 is provided with a fluorescent labeling substance probe, and the fluorescent labeling substance probe is an antibody 1 labeled by a fluorescent labeling substance.

The reaction detection spot 224 is immobilized with a reactive capture substance, which is a biotinylated antigen.

The reaction control point region 225 is immobilized with a biotinylated antibody 2-antigen complex.

The reaction detection point region 224 and the reaction quality control point region 225 are both bound with biotinylated antigen (or antibody or secondary antibody) and avidin which has undergone an amine-based condensation reaction with the lower sheet 22.

The fluorescent labeling substance is polystyrene microspheres which are purchased from Sammeishell science and technology Limited and have the diameter of 100-500nm, preferably 100-500 nm. Fluorescent materials are wrapped in the polystyrene microspheres, and the fluorescent materials are rhodamine, fluorescein isothiocyanate, phycoerythrin, lanthanide elements or quantum dots and the like. When the sample flows through the immobilized fluorescent labeling substance, the fluorescent labeling substance is easily dissolved and suspended in the sample.

The utility model discloses a working process does: the sample is purified by an affinity chromatography column 1 and then detected in a microfluidic chip 2.

When purifying the sample: the affinity chromatography column 1 can purify samples such as whole blood, serum, and plasma. Adding a sample to be purified into a binding buffer solution, wherein the binding buffer solution is a phosphate buffer solution with the pH value of 7, adding the binding buffer solution added with the sample into an affinity chromatography column 1, adding a binding buffer solution with the volume of 5-10 times that of curdlan after the liquid is drained, then adding an elution buffer solution, wherein the elution buffer solution is a phosphate buffer solution with the pH value of 2.7, inserting the lower part of a liquid discharge nozzle 16 of the affinity chromatography column 1 into a hollow guide column of a microfluidic chip 2, and detecting.

When the sample is detected: the purified sample is eluted to the sample neutralization area 221 of the microfluidic chip 2 by the affinity chromatography column 1, and the sample is neutralized by phosphate buffer solidified in the sample neutralization area 221. The sample then flows through the microstructure area 222 where it is mixed with phosphate buffer and then into the capillary microchannel 212. When the sample flows through the fluorescence labeling area 223, the fluorescence labeling antibody solidified in the fluorescence labeling area 223 is redissolved and flows to the reaction detection point area 224 together with the sample, and is captured by the antigen solidified in the reaction detection point area 224, the antibody not labeled by the captured fluorescence labeling substance and the antibody to be detected enter the reaction quality control point area 225, the antibody labeled by the fluorescence labeling substance is captured by the reaction quality control point area 225, and the rest of the liquid enters the waste liquid area 226.

Examples

An antibody detection system based on a microfluidic chip is used for joint detection of female infertility antibodies, as shown in fig. 2, the lower surface of an upper sheet 21 is provided with five upper detection units, fluorescent marker probes are cured in fluorescent marker regions 223 of the five upper detection units, and the fluorescent marker probes are detection compounds of polystyrene microsphere coupling antisperm antibody 1(AsAb), polystyrene microsphere coupling antisperm antibody 1(EmAb), polystyrene microsphere coupling antiscardial phospholipid antibody 1(AcAb), polystyrene microsphere coupling antisperm antibody 1(AoAb) and polystyrene microsphere coupling antiscorm antibody 1 (ToxAb). The reaction detection spot 224 has the corresponding immobilized anti-sperm antibody 2(AsAb), anti-endometrial antibody 2(EmAb), anti-cardiolipin antibody 2(AcAb), anti-ovarian antibody 2(AoAb), and anti-toxoplasma antibody 2(ToxAb), and a 1: 1, AsAb, EmAb, AcAb, AoAb and ToxA. The antibodies 1 and 2 are antibodies of antigens, but the recognition antigen sites are different, and can be understood as different antibodies which can combine the antigens, the antibodies 1 and 2 are purchased from Wuhan Toiran biotechnology limited, and the antigens are purchased from Shanghai collar tide organisms. The site of control of reactivity 225 has immobilized anti-sperm antibody 2(AsAb), anti-endometrial antibody 2(EmAb), anti-cardiolipin antibody 2(AcAb), anti-ovarian antibody 2(AoAb), and anti-Toxoplasma antibody 2(ToxAb), as well as complexes of the corresponding antigens.

The affinity chromatographic column 1 is filled with a protective solution. Before use, the protective solution in the affinity chromatography column 1 is drained, 12ml of binding buffer solution is added, the binding buffer solution is phosphate buffer solution with the pH value of 7, a sample is added into the binding buffer solution, the mixture is uniformly mixed and then added into the affinity chromatography column 1, and after liquid is dripped, 15ml of binding buffer solution is added, and after liquid is dripped, the liquid is dripped. At this time, the antibody is bound to the affinity column 1.

The affinity chromatography column 1 is inserted into a guide hollow column 2111 of the microfluidic chip 2, and 0.2ml of elution buffer solution is added, wherein the elution buffer solution is phosphate buffer solution with the pH value of 2.7. The eluted antibody enters the sample neutralization zone 221, the immobilized phosphate buffer in the sample neutralization zone 221 is redissolved to form a solution of the antibody to be detected, and the phosphate buffer neutralizes the antibody to a pH of 7.0. Driven by capillary force, the antibody solution to be detected reaches the fluorescent labeling area 223, the fluorescent label probe redissolves, the antibody solution to be detected and the fluorescent label probe flow through the reaction detection point area 224 together, and competitively binds with the antigen solidified in the reaction detection point area 224 to form an antibody structure labeled by avidin-biotin-antibody 2-antigen-antibody to be detected or avidin-biotin-antibody 2-antigen-fluorescent labeling substance. The unbound antibody to be detected and the fluorescent marker probe flow through the reactive mass control point region 225, and bind to the immobilized antigen in the reactive mass control point region 225, and the rest of the liquid enters the waste liquid region 226.

Through detection by a fluorescence detector, the fluorescence conjugates in the reaction detection point region 224 and the reaction quality control point region 225 are irradiated by light with a certain wavelength, and the detected fluorescence intensity is in opposite correlation with the substance to be detected.

Comparative example

An antibody detection system adopts an enzyme linked immunosorbent assay kit produced by Shenzhen Boka biotechnology limited, namely an antisperm antibody (AsAb) detection kit (enzyme linked immunosorbent assay), an anti-endometrium antibody (EmAb) detection kit (enzyme linked immunosorbent assay), an anti-cardiolipin antibody (AcAb) detection kit (enzyme linked immunosorbent assay), an anti-ovarian antibody (AoAb) detection kit (enzyme linked immunosorbent assay) and an anti-toxoplasma antibody (ToxAb) detection kit (enzyme linked immunosorbent assay).

Next, antibody detection tests were conducted by using examples and comparative examples, respectively.

Sample selection: selecting 40 positive samples and 20 negative samples of anti-sperm antibody (AsAb); 38 positive samples and 22 negative samples of anti-endometrial antibody (EmAb); 35 positive samples and 25 negative samples of anti-cardiolipin antibody (AcAb); anti-ovarian antibody (AoAb) positive samples 33, negative samples 27; 30 positive samples and 30 negative samples of anti-toxoplasma gondii antibody (ToxAb).

The above samples were subjected to the detection of antisperm antibody (AsAb), anti-endometrium antibody (EmAb), anti-cardiolipin antibody (AcAb), anti-ovarian antibody (AoAb) and anti-Toxoplasma antibody (ToxAb) using the antibody detection systems of examples and comparative examples, respectively, and the sensitivity was calculated according to the formula (1), and the specificity was calculated according to the formula (2), and the calculation results are shown in Table 1. As can be seen from Table 1, the sensitivity and specificity of the detection results of the antibody detection system of the example are superior to those of the antibody detection system of the comparative example.

Sensitivity is true positive number/(true positive number + false negative number) × 100% (1)

Specificity ═ number of true negatives/(number of true negatives + number of false positives) × 100% (2)

TABLE 1 comparative data sheet for antibody detection tests using examples and comparative examples, respectively

The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (9)

1. The utility model provides an antibody detecting system based on micro-fluidic chip which characterized in that: including affinity chromatographic column (1) and micro-fluidic chip (2), affinity chromatographic column (1) is protein G affinity chromatographic column or protein A affinity chromatographic column, micro-fluidic chip (2) include last piece (21) and lower piece (22), go up piece (21) closely the pressfitting in the upper surface of lower piece (22), sample adding hole (211) of lining up piece (21) are seted up in last piece (21), the bottom of affinity chromatographic column (1) is inserted in sample adding hole (211).

2. The microfluidic chip-based antibody detection system according to claim 1, wherein: the affinity chromatography column (1) comprises a column tube (11), a sample adding port (15) is formed in the top end of the column tube (11), a liquid discharging nozzle (16) is formed in the bottom end of the column tube (11), and a sand core filter sheet I (12), a filler (13) and a sand core filter sheet II (14) are sequentially arranged inside the column tube (11) from top to bottom.

3. The microfluidic chip-based antibody detection system according to claim 1, wherein: the lower surface of the upper piece (21) is provided with an upper detection unit, the upper detection unit comprises a capillary micro-channel (212) and a waste liquid groove (213), one end of the capillary micro-channel (212) is connected with the sample adding hole (211), and the other end of the capillary micro-channel is connected with the waste liquid groove (213).

4. The microfluidic chip-based antibody detection system according to claim 1, wherein: the lower surface of last piece (21) is equipped with two at least detection unit of going up, go up detection unit with the center of application of sample hole (211) distributes in as centre of a circle circumference in the outside of application of sample hole (211), it includes capillary microchannel (212) and waste liquid groove (213) to go up detection unit, the one end of capillary microchannel (212) with application of sample hole (211) are connected, the other end with waste liquid groove (213) are connected.

5. The microfluidic chip-based antibody detection system according to claim 3 or 4, wherein: the upper surface of lower piece (22) is just right the position department of application of sample hole (211) is equipped with sample neutralization area (221), the upper surface of lower piece (22) is just right last detecting element's position department is equipped with down detecting element, detecting element includes fluorescence mark district (223), reaction detection point district (224), reaction quality control point district (225), waste liquid district (226) that set gradually along the sample flow direction down, fluorescence mark district (223), reaction detection point district (224) and reaction quality control point district (225) are all just right capillary microchannel (212), waste liquid district (226) are just right waste liquid groove (213).

6. The microfluidic chip based antibody detection system of claim 5, wherein: the sample neutralizing area (221) is provided with a microstructure area (222) on the outer side, the microstructure area (222) is annular and is concentric with the sample neutralizing area (221), and a plurality of columnar or pyramid-shaped protrusions (2221) are uniformly distributed in the microstructure area (222) at intervals.

7. The microfluidic chip-based antibody detection system according to claim 1, wherein: the top edge of the sampling hole (211) vertically extends upwards to form a guide hollow column (2111), and an aluminum foil is sealed at the top of the guide hollow column (2111).

8. The microfluidic chip based antibody detection system of claim 5, wherein: the volumes of the capillary micro-channel (212) and the waste liquid groove (213) are both 5-300 mu L.

9. The microfluidic chip-based antibody detection system according to claim 1, wherein: the micro-fluidic chip (2) is made of quartz, glass or polyester substances and is circular or polygonal in shape.

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