Detection kit for galectin-3 binding protein

Abstract

The invention discloses a galectin-3 binding protein detection kit, which comprises a kit matrix disclosed by Chinese patent literature CN106771258A, and a hydroxyapatite agent accounting for 5-10% of the total amount of the kit matrix and a sodium alginate solution accounting for 4-7% of the total amount of the kit matrix are also added into the kit matrix. According to the detection kit disclosed by the invention, the kit matrix is adopted as a matrix raw material, and the hydroxyapatite agent and the sodium alginate solution are added and are matched, synergic and improved together, so that the obtained product has detection sensitivity, high detection precision and high detection efficiency.

Description

Technical Field

[0001] The present invention relates to the technical field of kits, and in particular to a kit for detecting galectin-3 binding protein. Background Art

[0002] Galectins are a family of sugar-binding proteins. All galectins have a common amino acid sequence in a domain called the carbohydrate recognition domain or CRD. Since the level of galectin-3 in body fluids such as blood or serum can reflect the disease state of the human body, such as chronic heart failure, accurate quantification of galectin-3 can be used to identify or predict diseases, to assess the severity of diseases, to stage or predict the consequences of diseases, or to monitor the effects of medications and other clinical or scientific research needs.

[0003] The existing test kit for detecting galectin-3 binding protein has poor detection efficiency and poor detection accuracy, resulting in insensitive detection and limiting detection efficiency. Based on this, the present invention further improves and processes it. Summary of the invention

[0004] In view of the defects of the prior art, the object of the present invention is to provide a detection kit for galectin-3 binding protein to solve the problems raised in the above background technology.

[0005] The present invention solves the technical problem by adopting the following technical solution: The present invention provides a galectin-3 binding protein detection kit, which comprises a kit matrix disclosed in Chinese patent document CN106771258A, wherein 5-10% of the total amount of the kit matrix is ​​added with a hydroxyapatite agent and 4-7% of the total amount of the kit matrix is ​​added with a sodium alginate solution.

[0006] Preferably, the preparation method of the hydroxyapatite agent is: S11: Heat-treat the hydroxyapatite at 75-85°C for 5-10 minutes, then place it in a sufficient amount of 5% by mass potassium permanganate solution and stir it evenly, then wash it with water and dry it; S12: adding 1-3 parts of silane coupling agent and 1-2 parts of urea to 4-7 parts of chitosan solution, and then adding 2-4 parts of sodium dodecylbenzene sulfonate, stirring and mixing to obtain a modified solution; S13: stirring and modifying the hydroxyapatite and modifying liquid of S11 at a weight ratio of 2:5, and after the stirring and modifying is completed, washing with water and drying to obtain a hydroxyapatite agent.

[0007] Preferably, the silane coupling agent is silane coupling agent KH560, and the mass fraction of the chitosan solution is 4-7%.

[0008] Preferably, the stirring speed of the stirring modification treatment is 450-500 r / min, and the stirring time is 35-45 min.

[0009] Preferably, the preparation method of the sodium alginate solution is: S101: performing a heat-homogenizing treatment on the carbon nanotubes to obtain a heat-homogenized carbon nanotube agent; S102: fully blending 3-5 parts of phosphate buffer solution, 1-3 parts of nano-silica sol, 2-5 parts of stearic acid and 3-5 parts of lanthanum chloride solution to obtain a regulator; S103: mixing the homogenized carbon nanotube agent and the regulator in a weight ratio of 4:1 and subjecting the mixture to ball milling at a speed of 1000-1500 r / min for 1-2 h. After the ball milling is completed, washing with water and drying are performed to obtain a regulated carbon nanotube agent; S104: The adjusted carbon nanotube agent and sodium alginate solution are mixed in a weight ratio of 2:5 to obtain sodium alginate.

[0010] Preferably, the specific operation steps of the heat treatment are: The carbon nanotubes are first heat treated at 130-140°C for 5-10 minutes, then heated to 245-255°C at a rate of 2-4°C / min, kept warm for 2-5 minutes, and finally cooled to 65°C at a rate of 1-3°C / min and kept warm.

[0011] Preferably, the pH value of the phosphate buffer solution is 5.0; and the mass fraction of the lanthanum chloride solution is 3-5%.

[0012] Preferably, the mass fraction of the sodium alginate solution is 6-10%.

[0013] Preferably, the preparation method of the detection kit comprises the following steps: fully mixing the kit matrix, hydroxyapatite agent and sodium alginate solution to obtain the detection kit.

[0014] Compared with the prior art, the present invention has the following beneficial effects: The detection kit of the present invention adopts a kit matrix as a matrix raw material, and by adding a hydroxyapatite agent and a sodium alginate solution, the two are coordinated and synergistically improved together, and the obtained product has detection sensitivity, high detection accuracy, and high detection efficiency. Since the hydroxyapatite in the hydroxyapatite agent is used as a point-like coordination system, the function of the matrix agent is enhanced, and at the same time, after being treated with a modified liquid, the coordinated synergistic effect of the hydroxyapatite agent and the sodium alginate solution is optimized. The sodium alginate solution adopts carbon nanotubes as a matrix, which enhances the compatibility of the raw material interface and the connectivity effect between the raw materials. At the same time, after being treated with homogenization and a regulating liquid for joint coordination, the performance effect of the product is further enhanced. DETAILED DESCRIPTION

[0015] The following is a clear and complete description of the technical solutions in the embodiments of the present invention in conjunction with specific embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0016] The present embodiment provides a galectin-3 binding protein detection kit, which includes the kit matrix disclosed in Chinese patent document CN106771258A, and the kit matrix further includes 5-10% of the total amount of the kit matrix and 4-7% of the sodium alginate solution.

[0017] The preparation method of the hydroxyapatite agent of this embodiment is: S11: Heat-treat the hydroxyapatite at 75-85°C for 5-10 minutes, then place it in a sufficient amount of 5% by mass potassium permanganate solution and stir it evenly, then wash it with water and dry it; S12: adding 1-3 parts of silane coupling agent and 1-2 parts of urea to 4-7 parts of chitosan solution, and then adding 2-4 parts of sodium dodecylbenzene sulfonate, stirring and mixing to obtain a modified solution; S13: stirring and modifying the hydroxyapatite and modifying liquid of S11 at a weight ratio of 2:5. After the stirring and modifying is completed, washing and drying are performed to obtain a hydroxyapatite agent.

[0018] The silane coupling agent of this embodiment is silane coupling agent KH560, and the mass fraction of the chitosan solution is 4-7%.

[0019] The stirring speed of the stirring modification treatment in this embodiment is 450-500 r / min, and the stirring time is 35-45 min.

[0020] The preparation method of the sodium alginate solution of this embodiment is: S101: performing a heat-homogenizing treatment on the carbon nanotubes to obtain a heat-homogenized carbon nanotube agent; S102: fully blending 3-5 parts of phosphate buffer solution, 1-3 parts of nano-silica sol, 2-5 parts of stearic acid and 3-5 parts of lanthanum chloride solution to obtain a regulator; S103: mixing the homogenized carbon nanotube agent and the regulator in a weight ratio of 4:1 and subjecting the mixture to ball milling at a speed of 1000-1500 r / min for 1-2 h. After the ball milling is completed, washing with water and drying are performed to obtain a regulated carbon nanotube agent; S104: The adjusted carbon nanotube agent and sodium alginate solution are mixed in a weight ratio of 2:5 to obtain sodium alginate.

[0021] The specific operation steps of the heat treatment in this embodiment are: The carbon nanotubes are first heat treated at 130-140°C for 5-10 minutes, then heated to 245-255°C at a rate of 2-4°C / min, kept warm for 2-5 minutes, and finally cooled to 65°C at a rate of 1-3°C / min and kept warm.

[0022] The pH value of the phosphate buffer solution of this embodiment is 5.0; the mass fraction of the lanthanum chloride solution is 3-5%.

[0023] The mass fraction of the sodium alginate solution in this embodiment is 6-10%.

[0024] The preparation method of the detection kit of this embodiment includes the following steps: the kit matrix, hydroxyapatite agent and sodium alginate solution are fully mixed to obtain the detection kit.

[0025] Example 1. The present embodiment provides a galectin-3 binding protein detection kit, which includes the kit matrix disclosed in Chinese patent document CN106771258A, and the kit matrix further includes 5% of the total amount of the kit matrix and 4% of the total amount of the kit matrix.

[0026] The preparation method of the hydroxyapatite agent of this embodiment is: S11: Heat-treat the hydroxyapatite at 75°C for 5 minutes, then place it in a sufficient amount of 5% by mass potassium permanganate solution and stir it evenly, then wash it with water and dry it; S12: adding 1 part of silane coupling agent and 1 part of urea to 4 parts of chitosan solution, and then adding 2 parts of sodium dodecylbenzene sulfonate, stirring and mixing to obtain a modified solution; S13: stirring and modifying the hydroxyapatite and modifying liquid of S11 at a weight ratio of 2:5, and after the stirring and modifying is completed, washing with water and drying to obtain a hydroxyapatite agent.

[0027] The silane coupling agent of this embodiment is silane coupling agent KH560, and the mass fraction of the chitosan solution is 4%.

[0028] The stirring speed of the stirring modification treatment in this embodiment is 450 r / min, and the stirring time is 35 min.

[0029] The preparation method of the sodium alginate solution of this embodiment is: S101: performing a heat-homogenizing treatment on the carbon nanotubes to obtain a heat-homogenized carbon nanotube agent; S102: fully blending 3 parts of phosphate buffer solution, 1 part of nano-silica sol, 2 parts of stearic acid and 3 parts of lanthanum chloride solution to obtain a regulator; S103: mixing the homogenized carbon nanotube agent and the regulator in a weight ratio of 4:1 and subjecting the mixture to ball milling at a speed of 1000-1500 r / min for 1-2 h. After the ball milling is completed, washing with water and drying are performed to obtain a regulated carbon nanotube agent; S104: The adjusted carbon nanotube agent and sodium alginate solution are mixed in a weight ratio of 2:5 to obtain sodium alginate.

[0030] The specific operation steps of the heat treatment in this embodiment are: The carbon nanotubes were first heat treated at 130°C for 5 min, then heated to 245°C at a rate of 2°C / min, kept at that temperature for 2 min, and finally cooled to 65°C at a rate of 1°C / min, and kept at that temperature.

[0031] The pH value of the phosphate buffer solution of this embodiment is 5.0; the mass fraction of the lanthanum chloride solution is 3-5%.

[0032] The mass fraction of the sodium alginate solution in this embodiment is 6%.

[0033] The preparation method of the detection kit of this embodiment comprises the following steps: fully mixing the kit matrix, hydroxyapatite agent and sodium alginate solution to obtain the detection kit.

[0034] Example 2. The present embodiment provides a galectin-3 binding protein detection kit, which includes the kit matrix disclosed in Chinese patent document CN106771258A, and the kit matrix further includes 10% of the total amount of the kit matrix of hydroxyapatite agent and 7% of the total amount of the kit matrix of sodium alginate solution.

[0035] The preparation method of the hydroxyapatite agent of this embodiment is: S11: Heat-treat the hydroxyapatite at 85°C for 10 minutes, then place it in a sufficient amount of 5% potassium permanganate solution and stir it evenly, then wash it with water and dry it; S12: adding 3 parts of silane coupling agent and 2 parts of urea to 7 parts of chitosan solution, and then adding 4 parts of sodium dodecylbenzene sulfonate, stirring and mixing to obtain a modified solution; S13: stirring and modifying the hydroxyapatite and modifying liquid of S11 at a weight ratio of 2:5. After the stirring and modifying is completed, washing and drying are performed to obtain a hydroxyapatite agent.

[0036] The silane coupling agent of this embodiment is silane coupling agent KH560, and the mass fraction of the chitosan solution is 4-7%.

[0037] The stirring speed of the stirring modification treatment in this embodiment is 500 r / min, and the stirring time is 45 min.

[0038] The preparation method of the sodium alginate solution of this embodiment is: S101: performing a heat-homogenizing treatment on the carbon nanotubes to obtain a heat-homogenized carbon nanotube agent; S102: fully blending 5 parts of phosphate buffer solution, 3 parts of nano-silica sol, 5 parts of stearic acid and 5 parts of lanthanum chloride solution to obtain a regulator; S103: mixing the homogenized carbon nanotube agent and the regulator in a weight ratio of 4:1 and subjecting the mixture to ball milling at a speed of 1500 r / min for 2 h. After the ball milling is completed, washing with water and drying are performed to obtain a regulated carbon nanotube agent; S104: The adjusted carbon nanotube agent and sodium alginate solution are mixed in a weight ratio of 2:5 to obtain sodium alginate.

[0039] The specific operation steps of the heat treatment in this embodiment are: The carbon nanotubes were first heat treated at 140°C for 10 min, then heated to 255°C at a rate of 4°C / min, kept at that temperature for 5 min, and finally cooled to 65°C at a rate of 3°C / min, and kept at that temperature.

[0040] The pH value of the phosphate buffer solution of this embodiment is 5.0; the mass fraction of the lanthanum chloride solution is 3-5%.

[0041] The mass fraction of the sodium alginate solution in this embodiment is 6-10%.

[0042] The preparation method of the detection kit of this embodiment comprises the following steps: fully mixing the kit matrix, hydroxyapatite agent and sodium alginate solution to obtain the detection kit.

[0043] Example 3. The present embodiment provides a galectin-3 binding protein detection kit, which includes the kit matrix disclosed in Chinese patent document CN106771258A, and the kit matrix further includes 7.5% of hydroxyapatite agent and 5.5% of sodium alginate solution in the total amount of the kit matrix.

[0044] The preparation method of the hydroxyapatite agent of this embodiment is: S11: Heat-treat the hydroxyapatite at 80°C for 7.5 minutes, then place it in a sufficient amount of 5% by mass potassium permanganate solution and stir it evenly, then wash it with water and dry it; S12: adding 2 parts of silane coupling agent and 1.5 parts of urea to 5.5 parts of chitosan solution, and then adding 3 parts of sodium dodecylbenzene sulfonate, stirring and mixing to obtain a modified solution; S13: stirring and modifying the hydroxyapatite and modifying liquid of S11 at a weight ratio of 2:5, and after the stirring and modifying is completed, washing with water and drying to obtain a hydroxyapatite agent.

[0045] The silane coupling agent of this embodiment is silane coupling agent KH560, and the mass fraction of the chitosan solution is 5.5%.

[0046] The stirring speed of the stirring modification treatment in this embodiment is 470 r / min, and the stirring time is 40 min.

[0047] The preparation method of the sodium alginate solution of this embodiment is: S101: performing a heat-homogenizing treatment on the carbon nanotubes to obtain a heat-homogenized carbon nanotube agent; S102: fully blending 4 parts of phosphate buffer solution, 2 parts of nano-silica sol, 3.5 parts of stearic acid and 4 parts of lanthanum chloride solution to obtain a regulator; S103: mixing the homogenized carbon nanotube agent and the regulator in a weight ratio of 4:1 and subjecting the mixture to ball milling at a ball milling speed of 1250 r / min for 1.5 h. After the ball milling is completed, washing with water and drying are performed to obtain a regulated carbon nanotube agent; S104: The adjusted carbon nanotube agent and sodium alginate solution are mixed in a weight ratio of 2:5 to obtain sodium alginate.

[0048] The specific operation steps of the heat treatment in this embodiment are: The carbon nanotubes were first heat treated at 135°C for 7.5 min, then heated to 250°C at a rate of 3°C / min, kept at that temperature for 3 min, and finally cooled to 65°C at a rate of 2°C / min, and kept at that temperature.

[0049] The pH value of the phosphate buffer solution in this embodiment is 5.0; the mass fraction of the lanthanum chloride solution is 4%.

[0050] The mass fraction of the sodium alginate solution in this embodiment is 8%.

[0051] The preparation method of the detection kit of this embodiment includes the following steps: the kit matrix, hydroxyapatite agent and sodium alginate solution are fully mixed to obtain the detection kit.

[0052] Comparative Example 1. The difference from Example 3 is that no hydroxyapatite agent was added.

[0053] Comparative Example 2. The difference from Example 3 is that no modifying liquid is added during the preparation of the hydroxyapatite agent.

[0054] Comparative Example 3. The difference from Example 3 is that no silane coupling agent or urea is added to the modified liquid.

[0055] Comparative Example 4. The difference from Example 3 is that no sodium alginate solution is added.

[0056] Comparative Example 5. The difference from Example 3 is that the method for preparing the sodium alginate solution is different; no homogenization treatment is used.

[0057] Comparative Example 6. The difference from Example 3 is that no conditioning agent is used in the preparation of the sodium alginate solution.

[0058] Comparative Example 7. The difference from Example 3 is that no nano silica sol and stearic acid are added to the regulator.

[0059] The products of Examples 1-3 and Comparative Examples 1-7 were subjected to performance tests, and the performance tests were as follows:

[0060] It can be seen from comparative examples 1-7 and embodiments 1-3 that; The product of the invention has excellent sensitivity detection efficiency, and at the same time, the product still has excellent stability under high temperature conditions. When no hydroxyapatite agent and no sodium alginate solution are added to the product, the performance of the product has an obvious tendency to deteriorate. When the two are coordinated and work together, the performance effect of the product is most significant. When no modifying liquid is added to the preparation of the hydroxyapatite agent, no silane coupling agent and urea are added to the modifying liquid, and the preparation method of the sodium alginate solution is different, when no homogenization treatment is adopted, no regulating agent is used in the preparation of the sodium alginate solution, and no nano-silica sol and stearic acid are added to the regulating agent, the performance of the product has a tendency to deteriorate. Only when the product raw material obtained by the method of the invention is adopted, the performance effect of the product is most significant.

[0061] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

[0062] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A detection kit for galectin-3 binding protein, the detection kit comprising a kit matrix disclosed in Chinese patent document CN106771258A, characterized in that: The kit matrix is ​​also added with 5-10% of the total amount of the kit matrix and 4-7% of the sodium alginate solution.

2. A detection kit for galectin-3 binding protein according to claim 1, characterized in that: The preparation method of the hydroxyapatite agent is: S11: Heat-treat the hydroxyapatite at 75-85°C for 5-10 minutes, then place it in a sufficient amount of 5% potassium permanganate solution and stir it evenly, then wash it with water and dry it; S12: adding 1-3 parts of silane coupling agent and 1-2 parts of urea to 4-7 parts of chitosan solution, and then adding 2-4 parts of sodium dodecylbenzene sulfonate, stirring and mixing to obtain a modified solution; S13: stirring and modifying the hydroxyapatite and modifying liquid of S11 at a weight ratio of 2:

5. After the stirring and modifying is completed, washing and drying are performed to obtain a hydroxyapatite agent.

3. A detection kit for galectin-3 binding protein according to claim 3, characterized in that: The silane coupling agent is silane coupling agent KH560, and the mass fraction of the chitosan solution is 4-7%.

4. A detection kit for galectin-3 binding protein according to claim 3, characterized in that: The stirring speed of the stirring modification treatment is 450-500 r / min, and the stirring time is 35-45 min.

5. A detection kit for galectin-3 binding protein according to claim 3, characterized in that: The preparation method of the sodium alginate solution is: S101: performing a heat-homogenizing treatment on the carbon nanotubes to obtain a heat-homogenized carbon nanotube agent; S102: fully blending 3-5 parts of phosphate buffer solution, 1-3 parts of nano-silica sol, 2-5 parts of stearic acid and 3-5 parts of lanthanum chloride solution to obtain a regulator; S103: mixing the homogenized carbon nanotube agent and the regulator in a weight ratio of 4:1 and subjecting the mixture to ball milling at a speed of 1000-1500 r / min for 1-2 h. After the ball milling is completed, washing with water and drying are performed to obtain a regulated carbon nanotube agent; S104: The adjusted carbon nanotube agent and sodium alginate solution are mixed in a weight ratio of 2:5 to obtain sodium alginate.

6. A detection kit for galectin-3 binding protein according to claim 5, characterized in that: The specific operation steps of the heat treatment are: The carbon nanotubes are first heat treated at 130-140°C for 5-10 minutes, then heated to 245-255°C at a rate of 2-4°C / min, kept warm for 2-5 minutes, and finally cooled to 65°C at a rate of 1-3°C / min and kept warm.

7. A detection kit for galectin-3 binding protein according to claim 5, characterized in that: The pH value of the phosphate buffer solution is 5.0; the mass fraction of the lanthanum chloride solution is 3-5%.

8. A detection kit for galectin-3 binding protein according to claim 5, characterized in that: The mass fraction of the sodium alginate solution is 6-10%.

9. A detection kit for galectin-3 binding protein according to claim 1, characterized in that: The preparation method of the detection kit comprises the following steps: fully mixing the kit matrix, hydroxyapatite agent and sodium alginate solution to obtain the detection kit.

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