Tumor abnormal sugar chain detection test paper and preparation method thereof

Abstract

The present application relates to the technical field of tumor abnormal sugar chain detection, and particularly relates to a tumor abnormal sugar chain detection test paper and a preparation method thereof, which comprises the following steps: cutting nitrocellulose membrane as a test paper substrate, uniformly coating a detection complex solution on the test paper substrate, drying in a 37 DEG C oven for 2-3 hours, adding a standard solution with a known concentration at one end of the test paper as a quality control line, dividing a reaction area on the test paper, sealing in an aluminum foil bag, storing at 4 DEG C in the dark, and obtaining the tumor abnormal sugar chain detection test paper. The detection test paper prepared by the present application only needs to drop the sample in the designated reaction area, without complex operation steps or professional equipment, so that the use threshold is reduced, and rapid preliminary screening is facilitated. The user can observe the color change or agglutination reaction on the test paper by naked eyes, so as to rapidly judge whether the sample contains abnormal sugar chains, and the detection efficiency is improved.

Description

Technical Field

[0001] This invention relates to the field of tumor abnormal glycan detection technology, and in particular to a tumor abnormal glycan detection test strip and its preparation method. Background Technology

[0002] Tumor abnormal glycan detection strips are a tool for the early detection of tumor risk. A significant characteristic of tumor cell occurrence and development is the abnormal change in the glycan structure on the cell membrane surface, a process known as "abnormal glycosylation." Glycosylation is a crucial post-translational modification of proteins, affecting their function, stability, and localization. When normal cells become malignant, the activity of glycosylation-modifying enzymes (such as glycosyltransferases and glycosidases) changes significantly, leading to abnormal alterations in the cell surface glycan structure and the production of various abnormal glycan glycoproteins (TAPs). TAPs are a class of abnormal glycoproteins released into body fluids during tumor cell metabolism, possessing unique glycan structures that differ significantly from glycoproteins produced by normal cells. Numerous studies have demonstrated a close relationship between TAPs and the occurrence, development, metastasis, and prognosis of malignant tumors. Detecting the presence and concentration of TAPs in body fluids (such as blood) can provide important evidence for early tumor detection, auxiliary diagnosis, efficacy evaluation, and monitoring of recurrence and metastasis. The core principle of TAP detection technology is that when normal cells become malignant, the activity of glycosylation-modifying enzymes changes, the sugar structure on the cell membrane surface changes, thereby producing a variety of abnormal glycan glycoproteins, which are eventually released into the body fluids.

[0003] In existing technologies, because TAP is not a specific marker of tumor cells, it can also be produced in certain non-tumor conditions, such as autoimmune diseases, inflammation, and infection. Therefore, TAP test strips have a certain false positive rate, meaning they may misdiagnose non-tumor patients as tumor patients. Furthermore, some physiological or pathological conditions, such as pregnancy, diabetes, and active tuberculosis, can also affect TAP test results, leading to false positives or false negatives. In addition, the sensitivity of TAP test strips is affected by various factors, such as sample collection, storage conditions, and testing time. These factors can cause fluctuations in the sensitivity of the test strip, resulting in unstable test results and affecting the accuracy of diagnosis. Although TAP test strips have certain advantages in early tumor screening, their sensitivity may still be insufficient for some early, small tumor lesions, making accurate detection difficult. Summary of the Invention

[0004] To address the problems mentioned in the background section, this invention provides a test strip for detecting abnormal glycan chains in tumors and its preparation method.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: A method for preparing a tumor abnormal glycan detection test strip includes the following steps: Nitrocellulose membrane was cut to serve as the test strip base. The detection complex solution was evenly coated onto the test strip base and dried in a 37°C oven for 2-3 hours. TAP standard solutions with known concentrations of 100 ng / mL, 200 ng / mL, 500 ng / mL, and 1000 ng / mL were added to one end of the test strip as quality control lines. The reaction zone, i.e. the area where agglutination reaction occurs after sample addition, was marked on the test strip. The strip was then sealed in an aluminum foil bag and stored at 4°C in the dark to obtain the tumor abnormal glycan detection test strip. The detection complex solution includes a recognition element solution and a signal conversion element solution.

[0006] Furthermore, the detection of the complex solution includes the following steps: S1. Add the aptamer or glyco-binding protein to a phosphate buffer solution with a pH of 7.4, add N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, stir the reaction at room temperature for 2-3 hours, dialyze or centrifuge and filter to obtain the recognition element solution. S2. Under an oxygen-free and light-protected environment, silver nitrate is quickly added to a solution containing 30-40% ascorbic acid, sodium citrate is added, and the mixture is heated and stirred until the solution turns yellow or light gray. The solution is then cooled to room temperature to obtain a silver nanoparticle solution. S3. Dimethyl terephthalate, ethylene glycol, and tetrabutyl titanate are added to a reactor containing methanol solvent. Under nitrogen protection, the mixture is heated and stirred for 3-4 hours to obtain an oligomer. Polyethylene glycol 200 is then added, and the mixture is heated and stirred for 4-6 hours. After the reaction is complete, unreacted ethylene glycol is removed by vacuum distillation until no liquid distills out. While still hot, N,N-dimethyl-2-propyl-2-enoxyethanolamine and azobisisobutyronitrile are added. The reaction temperature is controlled, and the mixture is stirred for 24-48 hours. Toluene can be added as needed to maintain the stability of the solution volume and concentration to obtain a copolymer solution. The silver nanoparticle solution is uniformly dispersed in the copolymer solution and stirred for 1-2 hours to obtain a signal conversion element solution. S4. Mix the recognition element solution with the signal conversion element solution and incubate at room temperature for 1-2 hours to obtain the detection complex solution; Among them, the aptamers are single-stranded DNA or RNA molecules screened from random libraries using SELEX technology, which can bind to the target TAP with high specificity. The glycoprotein is a lectin with glycan recognition ability, including one of concanavalin A (ConA), wheat germ lectin (WGA), ricin B chain, datura lectin (DSL), or peanut lectin (PNA).

[0007] Furthermore, the area of ​​the test strip substrate is 5cm × 1cm, the standard solution is dropped into a 1cm × 0.5cm area of ​​the test strip, and the area of ​​the reaction zone is 2cm × 1cm.

[0008] Further, in step S1, the mass ratio of aptamer or glycobinding protein, phosphate buffer, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is (1-2):10:(0.55-0.57):(0.48-0.52).

[0009] Further, in step S2, the mass ratio of silver nitrate, sodium citrate, and ascorbic acid is (1.5-1.8):(2-3):(8-10).

[0010] Furthermore, in step S2, the heating temperature is 90-100℃, the stirring speed is 50-100rpm, and the reaction time is 20-30min.

[0011] Further, in step S3, the mass ratio of dimethyl terephthalate, ethylene glycol, tetrabutyl titanate, polyethylene glycol 200, N,N-dimethyl-2-propyl-2-enoxyethanolamine, and azobisisobutyronitrile is (2.5-3.3):(1-1.5):(0.1-0.3):(0.8-1):(0.7-0.8):(0.05-0.1), and the mass ratio of copolymer solution to silver nanoparticle solution is (1.8-2.2):(0.8-1).

[0012] Furthermore, in step S3, the heating temperature is 190-200℃, the temperature rise is 220-230℃, the reaction temperature is controlled at 80-100℃, and the stirring speed is 50-100 rpm.

[0013] Further, in step S4, the mass ratio of the identification element solution to the signal conversion element solution is (1-1.2):(0.7-0.8).

[0014] According to another aspect of the present invention, a tumor abnormal glycan detection test strip prepared by the above preparation method is provided.

[0015] The beneficial effects of this invention are: 1. In the technical solution of this invention, the aptamer or glyco-binding protein maintains its active conformation in a phosphate buffer solution with a pH of 7.4. After activation by N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, the carboxyl groups on the surface of the protein are converted into active esters, which can covalently link with amino groups on other molecules or carriers, ensuring accurate identification of target molecules during the detection process and improving the accuracy and effectiveness of the test strip.

[0016] 2. In the technical solution of this invention, a polyethylene terephthalate-polyethylene glycol block copolymer is obtained by copolymerizing dimethyl terephthalate with ethylene glycol and polyethylene glycol. The copolymer possesses hydrophilic polyethylene glycol segments and hydrophobic polyethylene terephthalate segments, enabling it to form stable micelle or vesicle structures in aqueous solutions. When silver nanoparticles are encapsulated within this copolymer, the polyethylene glycol segments prevent direct contact between the nanoparticles, effectively preventing aggregation and precipitation, and acting as a bridge between the recognition element and the silver nanoparticles. The polyethylene terephthalate segments provide excellent chemical stability, resisting the influence of the external environment on the performance of the silver nanoparticles. Furthermore, the introduction of N,N-dimethyl-2-propyl-2-enoxyethanolamine further enhances the chemical stability of the copolymer, allowing it to maintain structural integrity under various conditions.

[0017] 3. In the technical solution of this invention, during the binding process of the recognition element and the signal conversion element, the active ester on the protein surface covalently bonds with the amino groups on the silver nanoparticle / copolymer complex, forming a stable complex. The polyethylene glycol segments and N,N-dimethyl-2-propyl-2-enoxyethanolamine possess certain biocompatibility, which can reduce interactions with organisms, promote the interaction between the detection complex and biological cells, improve the efficiency and accuracy of recognition, reduce immunogenicity, and help reduce the toxic effects of the detection complex on cells.

[0018] 4. In the technical solution of this invention, the silver nanoparticles in the signal conversion element solution have an amplified optical effect, enabling sensitive detection even at very low target molecule concentrations through changes in the optical properties of the silver nanoparticles. By adjusting the surface plasmon resonance effect of the silver nanoparticles, the changes in their optical properties are enhanced, allowing the detection complex to generate more sensitive and obvious signal changes when recognizing abnormal glycan chains in tumors.

[0019] 5. In this invention, the sample only needs to be added to the designated reaction area, without complicated operating procedures or specialized equipment, thus lowering the barrier to entry and facilitating rapid preliminary screening. Users can visually observe color changes or agglutination reactions on the test strip, thereby quickly determining whether the sample contains abnormal sugar chains, improving detection efficiency. Detailed Implementation

[0020] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Unless otherwise specified, the raw materials used in this invention are all from commercially available conventional products.

[0022] Preparation Example 1 The solution for detecting the complex is prepared by the following steps: S1. Add 100 mg of concanavalin A to 1 g of phosphate buffer at pH 7.4, add 55 mg of N-hydroxysuccinimide and 48 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, stir and react at room temperature for 2 h, then centrifuge and filter to obtain the recognition element solution. S2. In an oxygen-free and light-protected environment, prepare a 30% ascorbic acid solution with 800 mg ascorbic acid and deionized water. Quickly add 150 mg silver nitrate, then add 200 mg sodium citrate. Heat to 90°C and stir at 50 rpm for 20 min until the solution turns light gray. Cool to room temperature to obtain a silver nanoparticle solution. S3. Add 250 mg of dimethyl terephthalate, 100 mg of ethylene glycol, and 10 mg of tetrabutyl titanate to a reactor containing 1 g of methanol solvent. Under nitrogen protection, heat to 190 °C and stir at 50 rpm for 3 h to obtain oligomers. Continue to add 80 mg of polyethylene glycol 200, raise the temperature to 220 °C, and stir at 50 rpm for 4 h. After the reaction is completed, remove unreacted ethylene glycol by vacuum distillation until no liquid distills out. While hot, add 70 mg of N,N-dimethyl-2-propyl-2-enoxyethanolamine and 5 mg of azobisisobutyronitrile. Control the reaction temperature at 80 °C and stir for 24 h to obtain a copolymer solution. Disperse 80 mg of silver nanoparticle solution uniformly in 180 mg of copolymer solution and stir for 1 h to obtain a signal conversion element solution. S4. Mix 100 mg of recognition element solution with 70 mg of signal conversion element solution and incubate at room temperature for 1 h to obtain the detection complex solution.

[0023] Preparation Example 2 The solution for detecting the complex is prepared by the following steps: S1. Add 150 mg of wheat germ lectin to 1 g of phosphate buffer at pH 7.4, add 56 mg of N-hydroxysuccinimide and 50 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, stir and react at room temperature for 2.5 h, dialyze or centrifuge and filter to obtain the recognition element solution. S2. In an oxygen-free and light-protected environment, prepare a 35% ascorbic acid solution with 900 mg ascorbic acid and deionized water. Quickly add 165 mg silver nitrate, then add 250 mg sodium citrate. Heat to 95°C and stir at 60 rpm for 25 min until the solution turns light gray. Cool to room temperature to obtain a silver nanoparticle solution. S3. Add 300 mg of dimethyl terephthalate, 125 mg of ethylene glycol, and 20 mg of tetrabutyl titanate to a reactor containing 1 g of methanol solvent. Under nitrogen protection, heat to 195 °C and stir at 60 rpm for 3.5 h to obtain oligomers. Continue to add 90 mg of polyethylene glycol 200, raise the temperature to 225 °C, and stir at 60 rpm for 5 h. After the reaction is completed, remove unreacted ethylene glycol by vacuum distillation until no liquid distills out. While hot, add 75 mg of N,N-dimethyl-2-propyl-2-enoxyethanolamine and 8 mg of azobisisobutyronitrile. Control the reaction temperature at 90 °C and stir for 36 h to obtain a copolymer solution. Disperse 90 mg of silver nanoparticle solution uniformly in 200 mg of copolymer solution and stir for 1.5 h to obtain a signal conversion element solution. S4. Mix 110 mg of recognition element solution with 75 mg of signal conversion element solution and incubate at room temperature for 1.5 h to obtain the detection complex solution.

[0024] Preparation Example 3 The solution for detecting the complex is prepared by the following steps: S1. Add 200 mg of datura lectin to 1 g of phosphate buffer at pH 7.4, add 57 mg of N-hydroxysuccinimide and 52 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, stir and react at room temperature for 3 h, dialyze or centrifuge and filter to obtain the recognition element solution. S2. In an oxygen-free and light-protected environment, prepare a 40% ascorbic acid solution with 1000 mg ascorbic acid and deionized water. Quickly add 180 mg silver nitrate, then add 300 mg sodium citrate. Heat to 100°C and stir at 100 rpm for 30 min until the solution turns light gray. Cool to room temperature to obtain a silver nanoparticle solution. S3. Add 330 mg of dimethyl terephthalate, 150 mg of ethylene glycol, and 30 mg of tetrabutyl titanate to a reactor containing 1 g of methanol solvent. Under nitrogen protection, heat to 200 °C and stir at 100 rpm for 4 h to obtain oligomers. Continue to add 100 mg of polyethylene glycol 200, raise the temperature to 230 °C, and stir at 100 rpm for 6 h. After the reaction is completed, remove unreacted ethylene glycol by vacuum distillation until no liquid distills out. While hot, add 80 mg of N,N-dimethyl-2-propyl-2-enoxyethanolamine and 10 mg of azobisisobutyronitrile. Control the reaction temperature at 100 °C and stir for 48 h to obtain a copolymer solution. Disperse 100 mg of silver nanoparticle solution uniformly in 220 mg of copolymer solution and stir for 2 h to obtain a signal conversion element solution. S4. Mix 120 mg of recognition element solution with 80 mg of signal conversion element solution and incubate at room temperature for 2 h to obtain the detection complex solution.

[0025] Example 1 A method for preparing a tumor abnormal glycan detection test strip includes the following steps: Nitrocellulose membrane was cut into 5cm × 1cm pieces to serve as the test strip substrate. The detection complex solution prepared in Preparation Example 1 was uniformly coated onto the test strip substrate and dried in a 37°C oven for 2 hours. TAP standard solutions with known concentrations of 100 ng / mL, 200 ng / mL, 500 ng / mL, and 1000 ng / mL were added dropwise to one end of the test strip (a 1cm × 0.5cm area) as a quality control line. A 1cm × 0.5cm reaction zone was marked on the test strip, which is the area where agglutination reaction occurs after sample addition. The strip was then sealed in an aluminum foil bag and stored at 4°C in the dark to obtain the tumor abnormal glycan detection test strip.

[0026] Example 2 A method for preparing a tumor abnormal glycan detection test strip includes the following steps: Nitrocellulose membrane was cut into 5cm × 1cm pieces to serve as the test strip substrate. The detection complex solution prepared in Preparation Example 2 was uniformly coated onto the test strip substrate and dried in a 37°C oven for 2.5h. TAP standard solutions with known concentrations of 100ng / mL, 200ng / mL, 500ng / mL, and 1000ng / mL were dropped onto one end of the test strip (a 1cm × 0.5cm area) as a quality control line. A 1cm × 0.5cm reaction zone was marked on the test strip, which is the area where agglutination reaction occurs after sample addition. The strip was then sealed in an aluminum foil bag and stored at 4°C in the dark to obtain the tumor abnormal glycan detection test strip.

[0027] Example 3 A method for preparing a tumor abnormal glycan detection test strip includes the following steps: Nitrocellulose membrane was cut into 5cm × 1cm pieces to serve as the test strip substrate. The detection complex solution prepared in Preparation Example 3 was uniformly coated onto the test strip substrate and dried in an oven at 37°C for 3 hours. TAP standard solutions with known concentrations of 100ng / mL, 200ng / mL, 500ng / mL, and 1000ng / mL were dropped onto one end of the test strip (a 1cm × 0.5cm area) as a quality control line. A 1cm × 0.5cm reaction zone was marked on the test strip, which is the area where agglutination reaction occurs after sample addition. The strip was then sealed in an aluminum foil bag and stored at 4°C in the dark to obtain the tumor abnormal glycan detection test strip.

[0028] Comparative Example 1 The difference between this comparative example and Preparation Example 1 is that N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide are not added; the remaining steps are the same as in Preparation Example 1.

[0029] Comparative Example 2 The difference between this comparative example and Preparation Example 2 is that dimethyl terephthalate is not added; the remaining steps are the same as in Preparation Example 2.

[0030] Comparative Example 3 The difference between this comparative example and Preparation Example 3 is that N,N-dimethyl-2-propyl-2-enoxyethanolamine is not added; the remaining steps are the same as in Preparation Example 3.

[0031] Comparative Example 4 The difference between this comparative example and Example 1 is that the detection complex solution prepared in Comparative Example 1 is used, while the remaining steps are the same as in Example 1.

[0032] Comparative Example 5 The difference between this comparative example and Example 2 is that the detection complex solution prepared in Comparative Example 2 is used, while the remaining steps are the same as in Example 2.

[0033] Comparative Example 6 The difference between this comparative example and Example 3 is that the detection complex solution prepared in Comparative Example 3 is used, while the remaining steps are the same as in Example 3.

[0034] (I) Sensitivity test: Weigh out TAP standard and prepare a series of TAP solutions of different concentrations using phosphate buffer (pH 7.4): 10 ng / mL, 20 ng / mL, 50 ng / mL, 100 ng / mL, 200 ng / mL, 500 ng / mL, and 1000 ng / mL. Using a micropipette, add 50 μL of TAP solution to the reaction area of ​​the test strips prepared in Examples 1-3 and Comparative Examples 4-6, respectively. Observe the color development of the test strips after 5 minutes. If a clear gray, light brown, or dark brown color change appears in the reaction area of ​​the test strip, it is recorded as a positive result. Record the number of positive results at each concentration. Repeat the experiment three times and calculate the positive rate. The results are shown in Table 1.

[0035]

[0036] (II) Specificity test: Five serum samples from healthy individuals (containing no TAP or at very low concentrations) were collected. 50 μL of each sample was pipetted and added to the reaction area of ​​the test strips prepared in Examples 1-3 and Comparative Examples 4-6, respectively. The color development results were observed after 5 minutes. The color development results for each sample were recorded, and the false positive rate was calculated as: False positive rate = (Number of color-developed samples / Total number of experimental results) × 100%. Fifty replicate experiments were performed on each example and comparative example. The results are shown in Table 2.

[0037]

[0038] As shown in Table 1, Examples 1-3 all exhibited a 100% positive rate within the TAP concentration range of 10 ng / mL to 1000 ng / mL, indicating that the test strips have extremely high sensitivity and can accurately identify and reflect low to high concentrations of TAP. Comparative Example 4 showed a lower positive rate when the TAP concentration was below 100 ng / mL, and Comparative Example 5 also showed a lower positive rate when the TAP concentration was below 200 ng / mL.

[0039] As shown in Table 2, the false positive rate of Examples 1-3 was 0% in 50 repeated experiments, indicating that the test strips have extremely high specificity and can accurately distinguish between serum samples from healthy individuals (containing no TAP or with very low levels) and samples containing TAP. The false positive rates of Comparative Examples 4-6 were higher than those of the Examples, possibly due to enhanced reaction of the test strips to non-specific substances.

[0040] In Examples 1-3, concanavalin A, wheat germ lectin, and datura lectin serve as recognition elements, initiating the detection process through specific binding to TAP. N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide may help stabilize the structure of the recognition element, improving its binding affinity to TAP and thus increasing sensitivity. The signal conversion element, composed of silver nanoparticles and copolymers, is responsible for converting the binding signal between the recognition element and TAP into a visible color change. The addition of dimethyl terephthalate, N,N-dimethyl-2-propyl-2-enoxyethanolamine, etc., may help form a stable copolymer structure, improving signal conversion efficiency and thus reducing the false positive rate.

[0041] In summary, the test strips prepared in Examples 1-3 have good sensitivity and specificity.

[0042] In the description of this specification, the terms "preparation example," "example," "various examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that example or preparation example, which are included in at least one example or preparation example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same example or preparation example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more examples or preparation examples.

[0043] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing a test strip for detecting abnormal glycan chains in tumors, characterized in that, It is prepared by the following steps: Nitrocellulose membrane was cut and used as the test strip base. The detection complex solution was evenly coated on the test strip base and dried in an oven at 37°C for 2-3 hours. A standard solution of known concentration was dropped onto one end of the test strip as a quality control line. The reaction zone was marked on the test strip. The test strip was then sealed in an aluminum foil bag and stored in the dark at 4°C to obtain the tumor abnormal glycan detection test strip. The detection complex solution includes a recognition element solution and a signal conversion element solution; The solution for detecting the complex is prepared by the following steps: S1. Add the glyco-binding protein to phosphate buffer, add N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, stir the reaction at room temperature for 2-3 hours, dialyze or centrifuge and filter to obtain the recognition element solution. S2. Under an oxygen-free and light-protected environment, silver nitrate is quickly added to a solution containing 30-40% ascorbic acid, sodium citrate is added, and the mixture is heated and stirred until the solution turns yellow or light gray. The solution is then cooled to room temperature to obtain a silver nanoparticle solution. S3. Dimethyl terephthalate, ethylene glycol, and tetrabutyl titanate are added to a reactor containing methanol solvent. Under nitrogen protection, the mixture is heated and stirred for 3-4 hours to obtain an oligomer. Polyethylene glycol 200 is then added, and the mixture is heated and stirred for 4-6 hours. After the reaction is complete, the mixture is distilled under reduced pressure. While still hot, N,N-dimethyl-2-propyl-2-enoxyethanolamine and azobisisobutyronitrile are added. The reaction temperature is controlled, and the mixture is stirred for 24-48 hours to obtain a copolymer solution. The silver nanoparticle solution is uniformly dispersed in the copolymer solution and stirred for 1-2 hours to obtain a signal conversion element solution. S4. Mix the recognition element solution with the signal conversion element solution and incubate at room temperature for 1-2 hours to obtain the detection complex solution.

2. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, The area of ​​the test strip substrate is 5cm × 1cm. The standard solution is dropped into a 1cm × 0.5cm area of ​​the test strip, and the area of ​​the reaction zone is 2cm × 1cm.

3. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, In step S1, the mass ratio of glycobinding protein, phosphate buffer, N-hydroxysuccinimide and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide is (1-2):10:(0.55-0.57):(0.48-0.52).

4. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, In step S2, the mass ratio of silver nitrate, sodium citrate, and ascorbic acid is (1.5-1.8):(2-3):(8-10).

5. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, In step S2, the heating temperature is 90-100℃, the stirring speed is 50-100rpm, and the reaction time is 20-30min.

6. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, In step S3, the mass ratio of dimethyl terephthalate, ethylene glycol, tetrabutyl titanate, polyethylene glycol 200, N,N-dimethyl-2-propyl-2-enoxyethanolamine, and azobisisobutyronitrile is (2.5-3.3):(1-1.5):(0.1-0.3):(0.8-1):(0.7-0.8):(0.05-0.1), and the mass ratio of copolymer solution to silver nanoparticle solution is (1.8-2.2):(0.8-1).

7. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, In step S3, the heating temperature is 190-200℃, the temperature rise is 220-230℃, the reaction temperature is controlled at 80-100℃, and the stirring speed is 50-100 rpm.

8. The method for preparing a tumor abnormal glycan detection test strip according to claim 1, characterized in that, In step S4, the mass ratio of the identification element solution to the signal conversion element solution is (1-1.2):(0.7-0.8).

9. A tumor abnormal glycan detection test strip prepared by the preparation method according to any one of claims 1-8.

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