Fluorescent quantum dot-based m-BPA test paper and preparation method thereof

Abstract

The invention relates to the technical field of drug concentration detection, in particular to m-BPA detection test paper based on fluorescent quantum dots and a preparation method of the m-BPA detection test paper. The m-BPA detection test paper comprises a bottom plate, a water absorption pad, an NC membrane, a combination pad and a sample pad, and the sample pad, the combination pad, the NC membrane and the water absorption pad are sequentially overlapped and pasted on the bottom plate in the chromatography direction; the combination pad is provided with a carbon quantum dot-antibody conjugate, and the carbon quantum dot-antibody conjugate is a carbon quantum dot modified by an anti-m-BPA monoclonal antibody; a quality control line and a detection line are sequentially arranged on the NC membrane from the sample pad to the water absorption pad, the detection line is coated with an m-BPA-BSA antigen, and the quality control line is coated with a goat anti-mouse IgG antibody. The invention solves the problems that in the prior art, a rapid detection tool for m-BPA is lacked, and whether the boron concentration reaches the standard or not cannot be confirmed in time in an operation.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of drug concentration detection, and particularly relates to an m-BPA detection test paper based on fluorescent quantum dots and a preparation method thereof. BACKGROUND

[0002] With the rapid development of cancer treatment technology, BNCT (Boron Neutron Capture Therapy) has been widely used in clinical medicine. The basic principle of BCNT is that when a neutron beam is used to irradiate a patient injected with a boron carrying agent, the neutron and 10 B(n, α) reaction occurs, producing 7 Li and 4 He particles with a range of about 5-8 μm, which can destroy the DNA double helix of cancer cells and kill cancer cells, achieving the purpose of precise radiotherapy. 7 Li and 4 He has the advantages of heavy ion irradiation, which can effectively kill cells by destroying the DNA double helix of cells. Boron neutron capture therapy can achieve binary targeted therapy by combining boron injection and neutron irradiation technology, and has good efficacy for brain glioma, melanoma and head and neck recurrent tumors.

[0003] However, in order to achieve precise treatment of tumors by BNCT, it is necessary to determine the distribution and concentration (≥20 μg / g) of boron-10 drugs in the treatment target area before treatment, in order to achieve efficient treatment of tumors and protect normal tissues. If the quantitative distribution information of boron-10 drugs is not obtained before BNCT treatment, the uncertainty of the treatment dose will be caused, which will affect the treatment effect and cause unnecessary damage to normal tissues. The existing boron concentration detection methods mainly rely on ICP-MS detection, PET imaging and electrochemical sensors. ICP-MS detection is the gold standard method, which analyzes the boron content in tumor tissue by inductively coupled plasma mass spectrometry, but requires sample pretreatment (digestion, purification), professional equipment and operators, and the detection period is as long as 2-4 hours; PET imaging is performed by 11 C / m-BPA or 18 F-FBPA for preoperative imaging prediction, but cannot reflect the local boron concentration of the tumor in real time (the boron distribution after administration may be heterogeneous); electrochemical sensors require complex electrode preparation, and the detection stability is greatly affected by the environment, and have not been clinically transformed.

[0004] In existing technologies, m-boron phenylalanine (m-BPA), as a novel intravenous boron drug for BNCT, has a water solubility (125 g / L) that is more than 100 times that of the clinically commonly used p-boron phenylalanine (4-BPA) (0.72 g / L). It can be used to prepare a drug solution without the need for solubilization, and its tumor targeting is comparable to 4-BPA. However, there is currently a lack of rapid detection tools for m-BPA, making it impossible to immediately confirm whether the boron concentration is adequate during surgery. This can lead to either "ineffective irradiation" (insufficient boron concentration) or "over-irradiation" (excessive boron concentration causing damage to normal tissues). Summary of the Invention

[0005] In view of this, the purpose of this invention is to provide an m-BPA test strip based on fluorescent quantum dots and its preparation method, so as to at least solve the problem that there is a lack of rapid detection tools for m-BPA in the prior art and that boron concentration cannot be confirmed in real time during the operation.

[0006] The following is a summary of this disclosure to provide a basic understanding of some aspects. This summary is not intended to identify key or important elements, nor is it intended to limit the implementation or any aspects of the claims. Furthermore, this summary provides a simplified overview of some aspects that can be described in more detail in other parts of this disclosure.

[0007] The present invention solves the above-mentioned technical problems through the following technical means:

[0008] In a first aspect, embodiments of the present invention provide an m-BPA test strip based on fluorescent quantum dots, comprising a base plate, an absorbent pad, an NC membrane, a conjugate pad, and a sample pad, wherein the sample pad, conjugate pad, NC membrane, and absorbent pad are sequentially overlapped and adhered to the base plate in the direction of chromatography; the conjugate pad has a carbon quantum dot-antibody conjugate, wherein the carbon quantum dot-antibody conjugate is carbon quantum dots modified with anti-m-BPA monoclonal antibody; the NC membrane has a control line and a detection line sequentially arranged from the sample pad direction to the absorbent pad direction, the detection line is coated with m-BPA-BSA antigen, and the control line is coated with goat anti-mouse IgG antibody.

[0009] Preferably, the detection line is coated with 1 μg / cm of m-BPA-BSA antigen, and the control line is coated with 1.5 μg / cm of goat anti-mouse IgG antibody.

[0010] Preferably, the detection line and the control line are arranged parallel to each other on the NC membrane, with the detection line close to the conjugation pad and the control line close to the absorbent pad.

[0011] Preferably, the detection line and the control line are arranged parallel to each other on the NC membrane, with the detection line close to the conjugation pad and the control line close to the absorbent pad.

[0012] Preferably, the conjugate pad comprises a glass fiber membrane and a carbon quantum dot-antibody conjugate coated on the glass fiber membrane, wherein the glass fiber membrane is treated by immersion in a 0.01 mol / L PBS buffer solution at pH 7.4.

[0013] Preferably, the modification density of the anti-m-BPA monoclonal antibody is 4.5–5.5 μg / cm³. 2 .

[0014] Preferably, the sample pad has a pore size of 15–25 μm and a thickness of 0.4–0.6 mm, and the sample pad is a glass fiber membrane pretreated by soaking in a 0.01 mol / L PBS buffer solution with pH=7.4.

[0015] Preferably, the line width of both the quality control line and the detection line is 1 mm.

[0016] Secondly, the present invention also provides a method for preparing m-BPA detection test strip based on fluorescent quantum dots, comprising the following steps:

[0017] The synthesis of quantum dots involved dissolving citric acid and ethylenediamine in deionized water, reacting at 160–190 °C for 3.5–4.5 h, and then dialysis for 2–4 days after cooling to obtain a carbon quantum dot solution.

[0018] Antibody modification: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide were added to the carbon quantum dot solution and activated at room temperature for 25-35 min. Then, anti-m-BPA monoclonal antibody was added and reacted in PBS solution at pH 6.8 at room temperature for 1.5-2.5 h. BSA solution was added for blocking for 0.8-1.5 h, and the solution was purified by centrifugation to obtain the carbon quantum dot-antibody conjugate solution.

[0019] For the conjugate pad treatment, the glass fiber membrane was soaked in 0.01 mol / L PBS buffer solution (pH=7.4). The carbon quantum dot-antibody conjugate solution was then uniformly sprayed onto the treated glass fiber membrane, dried, and sealed for later use.

[0020] The membrane was coated with NC membrane, and goat anti-mouse IgG antibody was sprayed at the control line and m-BPA-BSA antigen was sprayed at the test line. The membrane was then dried.

[0021] Assemble the test strips by attaching them to the base plate in the order of sample pad, conjugate pad, NC membrane, and absorbent pad, with adjacent layers overlapping by 2mm. Cut the strips into 3cm×1cm strips and seal them in a package.

[0022] Preferably, the mass ratio of the carbon quantum dots, the anti-m-BPA monoclonal antibody, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide is 1:0.5:10:6.

[0023] Preferably, the BSA solution has a mass concentration of 1%; the centrifugal purification is performed by centrifugation at 10,000 rpm for 8-12 minutes, and the precipitate is resuspended in PBS solution at pH 6.8.

[0024] One or more technical solutions provided in the embodiments of the present invention have at least the following technical effects or advantages:

[0025] (1) The detection efficiency is greatly improved, and the detection time is only 5 minutes, which is 98% shorter than ICP-MS (2-4h), meeting the needs of intraoperative real-time judgment.

[0026] (2) Excellent detection performance, linear range of 0.1-100μg / g (covering the clinical actual tumor boron concentration range of 10-80μg / g), correlation coefficient R²=0.99; accuracy ≥98% (compared with ICP-MS, n=50 tumor samples); detection limit of 0.08μg / g, which is lower than the treatment threshold and there is no risk of missed detection.

[0027] (3) The operation is extremely simple, requiring no sample pretreatment or professional equipment (only a portable fluorescence reader is needed), and medical staff can master the operation in 5 minutes;

[0028] (4) It is minimally invasive and low-cost, requiring only 5-10 mg of tumor tissue, and the test strips can be mass-produced;

[0029] (5) It has strong stability. After being stored at 4℃ in a sealed container for 6 months, the fluorescence intensity decays by ≤8%, and the detection accuracy remains ≥97%, making it suitable for clinical storage.

[0030] (6) Strong adaptability: Designed for the intravenous administration characteristics of m-BPA, the detection results are consistent with the distribution pattern of the drug in vivo. Attached Figure Description

[0031] Figure 1 This is a schematic diagram of the structure of an m-BPA test strip based on fluorescent quantum dots;

[0032] The components include: sample pad 1, conjugate pad 2, NC membrane 3, absorbent pad 4, base plate 5, detection line 6, and quality control line 7. Detailed Implementation

[0033] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0034] For experiments not specifically described in the examples, the procedures or conditions should be followed according to the conventional experimental procedures described in the literature in this field. Reagents or instruments whose manufacturers are not specified are all commercially available conventional reagent products.

[0035] This application addresses the shortcomings of existing m-BPA boron concentration detection methods, which are "time-consuming, difficult to operate, and costly." Combining this with the intraoperative detection requirements of BNCT (Biovascular Non-Tissue Procedure), it provides a fluorescent quantum dot-based m-BPA test strip. Based on "antigen-antibody specific binding" and "fluorescence quenching effect," the surface of carbon quantum dots (CQDs) is modified with m-BPA-specific antibodies. When the test strip comes into contact with a homogenate of tumor tissue containing m-BPA, m-BPA specifically binds to the antibody, causing the fluorescence intensity of the carbon quantum dots to be linearly quenched as the m-BPA concentration increases. The m-BPA concentration is quantified by the fluorescence intensity.

[0036] Specifically, such as Figure 1 As shown, the m-BPA test strip based on fluorescent quantum dots of this application includes a base plate 5, an absorbent pad 4, an NC membrane 3, a conjugate pad 2, and a sample pad 1. The sample pad 1, conjugate pad 2, NC membrane 3, and absorbent pad 4 are sequentially overlapped and pasted on the base plate 5 in the chromatographic direction. The conjugate pad 2 has a carbon quantum dot-antibody conjugate, which is a carbon quantum dot modified with an anti-m-BPA monoclonal antibody. The NC membrane 3 has a control line 7 and a detection line 6 arranged sequentially from the sample pad 1 direction to the absorbent pad 4 direction. The detection line 6 is coated with m-BPA-BSA antigen, and the control line 7 is coated with goat anti-mouse IgG antibody.

[0037] In the specific implementation, detection line 6 is coated with 1 μg / cm of m-BPA-BSA antigen, and control line 7 is coated with 1.5 μg / cm of goat anti-mouse IgG antibody. Detection line 6 and control line 7 are arranged parallel to each other on NC membrane 3, with detection line 6 close to conjugate pad 2 and control line 7 close to absorbent pad 4. Conjugate pad 2 includes a glass fiber membrane and a carbon quantum dot-antibody conjugate coated on the glass fiber membrane. The glass fiber membrane is treated by soaking in 0.01 mol / L PBS buffer solution at pH 7.4. The modification density of anti-m-BPA monoclonal antibody is 4.5–5.5 μg / cm. 2The sample pad 1 has a pore size of 15–25 μm and a thickness of 0.4–0.6 mm. The sample pad 1 is a glass fiber membrane pretreated by soaking in 0.01 mol / L PBS buffer solution with pH=7.4. The line width of the control line 7 and the detection line 6 is 1 mm.

[0038] Sample pad 1 is used to receive tumor tissue homogenate samples and filter impurities; binding pad 2 is used to release fluorescent probes and bind to m-BPA; detection line 6 is used to capture fluorescent probes that have not bound to m-BPA; control line 7 is used to verify the effectiveness of the test strip and whether the fluorescent probes migrate normally; absorbent pad 4 has an absorbency capacity of ≥1mL and a thickness of 1mm and is used to drive the chromatographic migration of the sample; base plate 5 is used to fix the structure of each layer.

[0039] The method for preparing the m-BPA detection test strip based on fluorescent quantum dots in this application includes the following steps:

[0040] The synthesis of quantum dots involved dissolving citric acid and ethylenediamine in deionized water, reacting at 160–190 °C for 3.5–4.5 h, and then dialysis for 2–4 days after cooling to obtain a carbon quantum dot solution.

[0041] Antibody modification: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide were added to the carbon quantum dot solution and activated at room temperature for 25-35 min. Then, anti-m-BPA monoclonal antibody was added and reacted in PBS solution at pH 6.8 at room temperature for 1.5-2.5 h. BSA solution was added for blocking for 0.8-1.5 h, and the solution was purified by centrifugation to obtain the carbon quantum dot-antibody conjugate solution.

[0042] Combined with pad 2 treatment, the glass fiber membrane was soaked in 0.01 mol / L PBS buffer solution with pH=7.4, and the carbon quantum dot-antibody conjugate solution was uniformly sprayed onto the treated glass fiber membrane, dried, and sealed for later use.

[0043] The membrane was coated with NC membrane 3. Goat anti-mouse IgG antibody was sprayed at control line 7 on NC membrane 3, and m-BPA-BSA antigen was sprayed at detection line 6. The membrane was then dried.

[0044] Assemble the test strips by attaching them to the base plate 5 in the following order: sample pad 1, conjugate pad 2, NC membrane 3, and absorbent pad 4. Overlap adjacent layers by 2 mm, cut into 3cm × 1cm test strips, and seal them in a package.

[0045] The mass ratio of carbon quantum dots, anti-m-BPA monoclonal antibody, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and N-hydroxysuccinimide was 1:0.5:10:6. The mass concentration of the BSA solution was 1%. The purification was carried out by centrifugation at 10,000 rpm for 8–12 min, and the precipitate was resuspended in PBS solution at pH 6.8.

[0046] The following examples 1-3 will provide a detailed description of the m-BPA test strip based on fluorescent quantum dots and its preparation method of this application:

[0047] Example 1

[0048] The preparation method of the fluorescent quantum dot-based m-BPA detection test strip in this embodiment is as follows:

[0049] For the synthesis of quantum dots, 2 g of citric acid and 0.5 mL of ethylenediamine were dissolved in 20 mL of deionized water and reacted at 180 °C for 4 h. After cooling, the mixture was purified by dialyzing (molecular weight cutoff 1000 Da) for 3 days to obtain a carbon quantum dot solution. The concentration of this carbon quantum dot solution was 1 mg / mL, and the fluorescence quantum yield was 82.5%.

[0050] For antibody modification, take 1 mL of carbon quantum dot solution, add 10 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 6 mg of N-hydroxysuccinimide, activate at room temperature for 30 min, then add 100 μg of anti-m-BPA monoclonal antibody, react in PBS solution at pH 6.8 at room temperature for 2 h, add 1% BSA solution for blocking for 1 h, centrifuge at 10000 rpm for 10 min to purify, resuspend the precipitate in 1 mL of PBS solution at pH 6.8 to obtain carbon quantum dot-antibody conjugate solution.

[0051] For the conjugate pad 2 treatment, the glass fiber membrane was immersed in 0.01 mol / L PBS buffer solution (pH=7.4) for 5 min, and the modification density of anti-m-BPA monoclonal antibody was 5 μg / cm³. 2 The carbon quantum dot-antibody conjugate solution was uniformly sprayed onto the treated glass fiber membrane, dried, and sealed for later use.

[0052] The NC membrane 3 was coated with 1 μg / cm of m-BPA-BSA antigen at the detection line 6 and 1.5 μg / cm of goat anti-mouse IgG antibody at the control line 7. Goat anti-mouse IgG antibody was sprayed onto the control line 7 of the NC membrane 3 to form a C line, and m-BPA-BSA antigen was sprayed onto the detection line 6 to form a T line. The line width of both the control line 7 and the detection line 6 was controlled to be 1 mm. The membrane was then dried at 37°C for 2 h to obtain the NC membrane 3.

[0053] For the preparation of sample pad 1, a glass fiber membrane with a pore size of 20 μm and a thickness of 0.5 mm was immersed in 0.01 mol / L PBS buffer solution with pH=7.4 for 5 min to obtain sample pad 1.

[0054] Assemble the test strips by attaching them to the base plate 5 in the following order: sample pad 1, conjugate pad 2, NC membrane 3, and absorbent pad 4. Overlap the adjacent layers by 2 mm, cut them into 3cm × 1cm test strips, and seal them in a package.

[0055] Example 2

[0056] The preparation method of the fluorescent quantum dot-based m-BPA detection test strip in this embodiment is as follows:

[0057] For the synthesis of quantum dots, 2 g of citric acid and 0.5 mL of ethylenediamine were dissolved in 20 mL of deionized water and reacted at 160 °C for 4.5 h. After cooling, the mixture was purified by dialyzing (molecular weight cutoff 1000 Da) for 2 days to obtain a carbon quantum dot solution. The concentration of this carbon quantum dot solution was 1 mg / mL, and the fluorescence quantum yield was 82.3%.

[0058] For antibody modification, 1 mL of carbon quantum dot solution was taken, and 10 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 6 mg of N-hydroxysuccinimide were added. The mixture was activated at room temperature for 25 min, and then 100 μg of anti-m-BPA monoclonal antibody was added. The mixture was reacted in PBS solution at pH 6.8 at room temperature for 1.5 h. After blocking with 1% BSA solution for 0.8 h, the mixture was purified by centrifugation at 10000 rpm for 8 min. The precipitate was resuspended in 1 mL of PBS solution at pH 6.8 to obtain the carbon quantum dot-antibody conjugate solution.

[0059] For the conjugate pad 2 treatment, the glass fiber membrane was immersed in 0.01 mol / L PBS buffer solution (pH=7.4) for 5 min, with the anti-m-BPA monoclonal antibody modification density at 4.5 μg / cm³. 2 The carbon quantum dot-antibody conjugate solution was uniformly sprayed onto the treated glass fiber membrane, dried, and sealed for later use.

[0060] The NC membrane 3 was coated with 1 μg / cm of m-BPA-BSA antigen at the detection line 6 and 1.5 μg / cm of goat anti-mouse IgG antibody at the control line 7. Goat anti-mouse IgG antibody was sprayed onto the control line 7 of the NC membrane 3 to form a C line, and m-BPA-BSA antigen was sprayed onto the detection line 6 to form a T line. The line width of both the control line 7 and the detection line 6 was controlled to be 1 mm. The membrane was then dried at 37°C for 2 h to obtain the NC membrane 3.

[0061] For the preparation of sample pad 1, a glass fiber membrane with a pore size of 15 μm and a thickness of 0.4 mm was soaked in 0.01 mol / L PBS buffer solution with pH=7.4 for 5 min to obtain sample pad 1.

[0062] Assemble the test strips by attaching them to the base plate 5 in the following order: sample pad 1, conjugate pad 2, NC membrane 3, and absorbent pad 4. Overlap the adjacent layers by 2 mm, cut them into 3cm × 1cm test strips, and seal them in a package.

[0063] Example 3

[0064] The preparation method of the fluorescent quantum dot-based m-BPA detection test strip in this embodiment is as follows:

[0065] For the synthesis of quantum dots, 2 g of citric acid and 0.5 mL of ethylenediamine were dissolved in 20 mL of deionized water and reacted at 190 °C for 3.5 h. After cooling, the mixture was purified by dialyzing (molecular weight cutoff 1000 Da) for 4 days to obtain a carbon quantum dot solution. The concentration of this carbon quantum dot solution was 1 mg / mL, and the fluorescence quantum yield was 81.9%.

[0066] For antibody modification, take 1 mL of carbon quantum dot solution, add 10 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 6 mg of N-hydroxysuccinimide, activate at room temperature for 35 min, then add 100 μg of anti-m-BPA monoclonal antibody, react in PBS solution at pH=6.8 at room temperature for 2.5 h, add 1% BSA solution for blocking for 1.5 h, centrifuge at 10000 rpm for 12 min to purify, and resuspend the precipitate in 1 mL of PBS solution at pH=6.8 to obtain carbon quantum dot-antibody conjugate solution.

[0067] For the conjugate pad 2 treatment, the glass fiber membrane was immersed in 0.01 mol / L PBS buffer solution (pH=7.4) for 5 min, with the anti-m-BPA monoclonal antibody modification density at 5.5 μg / cm³. 2 The carbon quantum dot-antibody conjugate solution was uniformly sprayed onto the treated glass fiber membrane, dried, and sealed for later use.

[0068] The NC membrane 3 was coated with 1 μg / cm of m-BPA-BSA antigen at the detection line 6 and 1.5 μg / cm of goat anti-mouse IgG antibody at the control line 7. Goat anti-mouse IgG antibody was sprayed onto the control line 7 of the NC membrane 3 to form a C line, and m-BPA-BSA antigen was sprayed onto the detection line 6 to form a T line. The line width of both the control line 7 and the detection line 6 was controlled to be 1 mm. The membrane was then dried at 37°C for 2 h to obtain the NC membrane 3.

[0069] For the preparation of sample pad 1, a glass fiber membrane with a pore size of 25 μm and a thickness of 0.6 mm was soaked in 0.01 mol / L PBS buffer solution with pH=7.4 for 5 min to obtain sample pad 1.

[0070] Assemble the test strips by attaching them to the base plate 5 in the following order: sample pad 1, conjugate pad 2, NC membrane 3, and absorbent pad 4. Overlap the adjacent layers by 2 mm, cut them into 3cm × 1cm test strips, and seal them in a package.

[0071] Using the m-BPA test paper prepared in Example 1 as a sample, the following performance tests were performed:

[0072] (1) Test strip performance verification

[0073] ① Plot a standard curve. Prepare m-BPA standard solutions (0.1, 1, 10, 20, 50, 80, 100 μg / g) respectively. Take 50 μL of each solution and add it to the test paper. After 5 min, read the fluorescence intensity using a portable fluorescence reader and plot the standard curve: y = -0.032x + 8.56 (y is the fluorescence intensity, x is the m-BPA concentration), R² = 0.99.

[0074] ② Actual samples were tested. Tumor tissue (n=10) was collected from B16F10 tumor-bearing mice 60 min after intravenous injection of m-BPA (dose: 1 mg / 100 μL PBS, pH=7.4). The fluorescence intensity was read using the m-BPA test paper prepared in Example 1 and compared with the results of ICP-MS detection. The results are shown in Table 1.

[0075] The compatibility parameters between the portable fluorescence reader and the m-BPA test strip are shown in Tables 2 and 3.

[0076] Table 1

[0077]

[0078] Table 2

[0079]

[0080] Table 3

[0081]

[0082] As can be seen from the data in Table 1, the m-BPA test strip prepared in Example 1 showed no statistically significant difference from the ICP-MS detection (p>0.05), with a relative error ≤±3%, which meets clinical requirements.

[0083] (2) Anti-interference verification

[0084] When interfering substances such as serum, tumor tissue homogenate, 4-BPA, and o-BPA are added to the m-BPA standard solution (20 μg / g), the test strip response is as follows:

[0085] ① Serum / tissue homogenate interference: relative error ≤ ±2.1%;

[0086] ② 4-BPA / o-BPA cross-reactivity: relative error ≤ ±1.5%;

[0087] Therefore, the m-BPA test strip prepared in Example 1 has strong anti-interference properties and can be used for intraoperative complex sample detection.

[0088] The foregoing description includes examples from this specification. Of course, for the purposes of describing this specification, it is impossible to describe every conceivable combination of components or methods; however, those skilled in the art will understand that many other combinations and arrangements are possible. Therefore, this specification is intended to cover all such changes, modifications, and variations that fall within the spirit and scope of the appended claims. Furthermore, with regard to the use of the term "comprising" in the detailed description or claims, the term is intended to be inclusive in a manner similar to the term "including," as interpreted when "comprising" is used as a transitional word in the claims.

[0089] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A fluorescent quantum dot-based m-BPA test strip, comprising a base plate, characterized in that, The test paper also comprises a water absorption pad, an NC membrane, a combination pad and a sample pad, which are sequentially and adhesively connected on the base plate in the chromatographic direction; the combination pad is provided with carbon quantum dot-antibody conjugates, which are carbon quantum dots modified by anti-m-BPA monoclonal antibodies; the NC membrane is sequentially provided with a quality control line and a detection line from the sample pad to the water absorption pad, the detection line is coated with m-BPA-BSA antigen, and the quality control line is coated with sheep anti-mouse IgG antibody.

2. A fluorescent quantum dot-based m-BPA test strip according to claim 1, wherein, The detection line is coated with 1 μg / cm of m-BPA-BSA antigen, and the quality control line is coated with 1.5 μg / cm of sheep anti-mouse IgG antibody.

3. A fluorescent quantum dot-based m-BPA test strip according to claim 2, wherein, The detection line and the quality control line are arranged in parallel on the NC membrane, the detection line is close to the combination pad, and the quality control line is close to the water absorption pad.

4. The m-BPA test strip based on fluorescent quantum dots according to claim 1, wherein, The combination pad comprises a glass fiber membrane and carbon quantum dot-antibody conjugates coated on the glass fiber membrane, and the glass fiber membrane is soaked and treated by 0.01 mol / L PBS buffer solution with pH=7.

4.

5. A fluorescent quantum dot-based m-BPA test strip according to claim 4, wherein, The modification density of the anti-m-BPA monoclonal antibody is 4.5-5.5 μg / cm 2 .

6. A fluorescent quantum dot-based m-BPA test strip according to claim 1, wherein, The sample pad has a pore size of 15-25 μm and a thickness of 0.4-0.6 mm, and the sample pad is a glass fiber membrane soaked and pretreated by 0.01 mol / L PBS buffer solution with pH=7.

4.

7. A fluorescent quantum dot-based m-BPA test strip according to claim 1, wherein, The line width of the quality control line and the detection line is 1 mm.

8. A method for preparing a fluorescent quantum dot-based m-BPA test strip, characterized in that, The test paper comprises the following steps: Synthesis of quantum dots: citric acid and ethylenediamine are dissolved in deionized water, reacted at 160-190°C for 3.5-4.5 h, and then dialyzed for 2-4 days after cooling to obtain a carbon quantum dot solution; Antibody modification: 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide are added to the carbon quantum dot solution, activated at room temperature for 25-35 min, then anti-m-BPA monoclonal antibody is added, reacted at room temperature in PBS solution with pH=6.8 for 1.5-2.5 h, BSA solution is added for blocking for 0.8-1.5 h, and then centrifuged and purified to obtain a carbon quantum dot-antibody conjugate solution; Combination pad treatment: the glass fiber membrane is soaked and treated by 0.01 mol / L PBS buffer solution with pH=7.4, and the carbon quantum dot-antibody conjugate solution is uniformly sprayed on the treated glass fiber membrane, dried and sealed for standby; NC membrane coating: sheep anti-mouse IgG antibody is sprayed on the quality control line of the NC membrane, and m-BPA-BSA antigen is sprayed on the detection line, and then dried; Test paper assembly: the sample pad, the combination pad, the NC membrane and the water absorption pad are sequentially adhered on the base plate, the adjacent layers are overlapped by 2 mm, and then cut into test paper strips with a size of 3 cm×1 cm, and finally sealed and packaged.

9. The method of making according to claim 8, wherein, The mass ratio of the carbon quantum dots, the anti-m-BPA monoclonal antibody, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide is 1:0.5:10:

6.

10. The method of claim 8, wherein, The mass concentration of the BSA solution is 1%, and the centrifugal purification is centrifuged at a speed of 10000 rpm for 8-12 min, and the precipitate is resuspended in PBS solution with pH=6.8.

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