Anti-swelling enzyme-containing hydrogel and preparation method and application thereof

Abstract

The application provides an anti-swelling enzyme-containing hydrogel and a preparation method and application thereof, and constructs the enzyme-containing hydrogel by using a high-biocompatibility and super-hydrophilic sulfobetaine zwitterionic gel, so that water molecules can be pulled away from the hydrophobic region of the enzyme, and the activity of the enzyme is maximally maintained, and the interaction between glucose and the enzyme binding site is promoted, so that the enzyme deactivation is slowed down, and the service life of the sensor is prolonged; the pore size of the hydrogel is adjusted by using carboxylated cellulose nanofibers, so that the anti-swelling performance of the hydrogel is improved, the anti-swelling hydrogel has a higher liquid carrying capacity and a stable water retention rate, and the service life can be prolonged; and the stability and catalytic activity of the enzyme are increased by using platinum nanoparticles, and the electron mobility of the hydrogel is improved, so that the sensitivity of the sensor is improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of functional polymer hydrogel, in particular to an anti-swelling enzyme-containing hydrogel and a preparation method and application thereof. BACKGROUND

[0002] Diabetes is a world chronic disease that threatens human health and life, and blood glucose monitoring is the premise and basis for the diagnosis and treatment of diabetes, so the electrochemical glucose enzyme sensor has always been a special focus of electrochemical sensors. In recent years, the market demand for multifunctional hydrogel has been growing, and enzyme-containing hydrogel is increasingly used in electrochemical enzyme sensors. However, the traditional enzyme-containing hydrogel is prone to swelling, enzyme leakage, enzyme inactivation, and low liquid holding capacity, resulting in short service life. Therefore, it is necessary to continue to develop an anti-swelling enzyme-containing hydrogel with long service life. SUMMARY

[0003] The technical problem to be solved by the present application is to provide an anti-swelling enzyme-containing hydrogel.

[0004] Another technical problem to be solved by the present application is to provide a preparation method of the anti-swelling enzyme-containing hydrogel.

[0005] Another technical problem to be solved by the present application is to provide an application of the anti-swelling enzyme-containing hydrogel.

[0006] The technical solution adopted by the present application is:

[0007] An anti-swelling enzyme-containing hydrogel is prepared by mixing sulfobetaine as a monomer, methylene bisacrylamide as a crosslinking agent, ammonium persulfate as an initiator, and carboxylated cellulose nanofiber; platinum nanoparticles are added to increase the stability and catalytic activity of the enzyme; and the mixed solution is mixed with an enzyme solution and solidified in a water bath at 30-37 DEG C to obtain a hydrogel enzyme film.

[0008] The preparation method of the anti-swelling enzyme-containing hydrogel is as follows:

[0009] (1) Mix sulfobetaine monomer, carboxylated cellulose nanofiber, methylene bisacrylamide, ammonium persulfate, and PBS buffer solution, and then add platinum nanoparticle solution, stir to obtain a hydrogel precursor solution;

[0010] (2) Mix and stir the enzyme freeze-dried powder with PBS buffer solution to obtain a glucose oxidase solution;

[0011] (3) Mix the hydrogel precursor solution and the glucose oxidase solution in a volume ratio of 1:1 and solidify in a water bath at 37 DEG C for 24 hours to obtain an anti-swelling enzyme-containing hydrogel with long service life.

[0012] Preferably, in the preparation method of the anti-swelling enzyme-containing hydrogel, the mass-volume ratio of the sulfobetaine monomer, methylene bisacrylamide, ammonium persulfate, carboxylated cellulose nanofiber, PBS buffer and platinum nanoparticle solution in step (1) is (0.3-0.5) g:(4-6) mg:(2-4) mg:(0.3-0.5) ml:(0.1-0.2) ml:(70-80) μl.

[0013] Preferably, in the preparation method of the anti-swelling enzyme-containing hydrogel, the enzyme freeze-dried powder in step (2) is glucose oxidase, glucose dehydrogenase or catalase.

[0014] Preferably, in the preparation method of the anti-swelling enzyme-containing hydrogel, the solidification environment in step (3) should be at 30-37℃.

[0015] The proportions of the above mixtures can be increased or reduced in proportion.

[0016] The anti-swelling enzyme-containing hydrogel is used in the preparation of a diabetes detection device.

[0017] Preferably, in the use of the anti-swelling enzyme-containing hydrogel, the diabetes detection device is an electrochemical glucose enzyme sensor.

[0018] The anti-swelling enzyme-containing hydrogel has the following beneficial effects:

[0019] The anti-swelling enzyme-containing hydrogel uses a sulfobetaine zwitterionic gel with high biocompatibility and superhydrophilicity to construct an enzyme-containing hydrogel, so that water molecules can be pulled away from the hydrophobic region of the enzyme, thereby maintaining the activity of the enzyme to the greatest extent, and also promoting the interaction between glucose and the enzyme binding site, thereby slowing down the deactivation of the enzyme and prolonging the service life of the sensor; the carboxylated cellulose nanofiber is used to adjust the pore size of the hydrogel to improve the anti-swelling performance of the hydrogel, and the anti-swelling hydrogel has a high liquid holding capacity and a stable water retention rate, which can prolong the service life; the platinum nanoparticles are used to increase the stability and catalytic activity of the enzyme and improve the electron mobility of the hydrogel, thereby improving the sensitivity of the sensor; the preparation method is simple and easy to implement, the sulfobetaine zwitterionic hydrogel prepared has good biocompatibility with the enzyme, can pull water molecules away from the hydrophobic region of the enzyme, slows down the deactivation of the enzyme, and maintains the activity of the enzyme; has good anti-swelling performance, avoids enzyme leakage; has a high liquid holding capacity and a stable water retention rate, which can prolong the service life and is conducive to subsequent electrochemical measurement. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The anti-swelling enzyme-containing hydrogel provided by the application has the structure shown in the figure. DETAILED DESCRIPTION

[0021] To further illustrate the present application, the following examples are provided.

[0022] Example 1

[0023] A swelling-resistant enzyme-containing hydrogel, the preparation method thereof comprises the following steps:

[0024] (1) 0.343 g of sulfobetaine monomer, 400 μL of carboxylated cellulose nanofiber suspension (6 wt%), 5 mg of methylene bisacrylamide, 3 mg of ammonium persulfate, and 100 μl of PBS buffer were mixed, and then 75 μl of platinum nanoparticle solution (10%) was added. After stirring, a hydrogel precursor solution was obtained;

[0025] (2) Glucose oxidase freeze-dried powder (10000 v / g) was added to 500 μl of PBS buffer, mixed and stirred to obtain a glucose oxidase solution with a concentration of 100 mg / ml;

[0026] (3) Each of the above hydrogel precursor solution and the glucose oxidase solution 100 μl was mixed and solidified in a 37°C water bath for 24h to obtain a swelling-resistant long service life enzyme-containing hydrogel. The structure of the swelling-resistant enzyme-containing hydrogel is shown in Figure 1 .

[0027] Example 2

[0028] A swelling-resistant long service life enzyme-containing hydrogel preparation method, the specific steps are as follows:

[0029] (1) 0.343 g of sulfobetaine monomer, 400 μL of carboxylated cellulose nanofiber suspension (6 wt%), 5 mg of methylene bisacrylamide, 3 mg of ammonium persulfate, and 100 μl of PBS buffer were mixed, and then 75 μl of platinum nanoparticle solution (10%) was added. After stirring, a hydrogel precursor solution was obtained;

[0030] (2) Glucose dehydrogenase freeze-dried powder (10 mg) was added to 1 ml of PBS buffer, mixed and stirred to obtain a glucose oxidase solution with a concentration of 10 mg / ml;

[0031] (3) Each of the above hydrogel precursor solution and the glucose dehydrogenase solution 100 μl was mixed and solidified in a 37°C water bath for 24h to obtain a swelling-resistant long service life enzyme-containing hydrogel.

[0032] Example 3

[0033] The application of the swelling-resistant enzyme-containing hydrogel described in Example 1 in the preparation of an electrochemical glucose enzyme sensor.

[0034] (1) The anti-swelling enzyme-containing hydrogel described in Example 1 is solidified on the surface of the sensor electrode to form an electrochemical glucose enzyme sensor attached to the inner surface of the skin of the arm;

[0035] (2) The sensor extraction electrode is connected to a constant current source to extract the interstitial fluid through the skin and store it in the enzyme-containing hydrogel, and the extraction process lasts for 3 min;

[0036] (3) The sensor reaction electrode is connected to an electrochemical workstation, and the current signal generated by the reaction between the glucose oxidase in the hydrogel and the glucose in the interstitial fluid is used to characterize the glucose concentration in the current detection, and the measurement process lasts for 1 min;

[0037] (4) Repeat the above steps (2) and (3) to realize real-time continuous monitoring of blood glucose.

[0038] The above-described embodiments are only descriptions of the preferred embodiments of the present application and do not limit the scope of the present application. Without departing from the design spirit of the present application, various modifications and improvements to the technical solutions of the present application made by ordinary engineering technicians in the art shall fall within the protection scope determined by the claims of the present application.

Claims

1. An anti-swelling enzyme-containing hydrogel, characterized by: The sulfobetaine monomer is mixed with carboxylated cellulose nanofiber by using methylene bisacrylamide as a crosslinking agent and ammonium persulfate as an initiator; platinum nanoparticles are added to increase the stability and catalytic activity of the enzyme; the mixed solution is mixed with an enzyme solution and solidified in a 30℃-37℃ water bath to obtain a hydrogel enzyme film.

2. The method of preparing an anti-swell, enzyme-containing hydrogel of claim 1, characterized by: The specific steps are as follows: (1) The sulfobetaine monomer, carboxylated cellulose nanofiber, methylene bisacrylamide, ammonium persulfate, and PBS buffer solution are mixed, and then a platinum nanoparticle solution is added, and after stirring, a hydrogel precursor solution is obtained; (2) The enzyme lyophilized powder is added to the PBS buffer solution, mixed and stirred to obtain a glucose oxidase solution; (3) The above hydrogel precursor solution and glucose oxidase solution are mixed in a volume ratio of 1:1 and solidified in a 37℃ water bath for 24h to obtain an anti-swelling long-service-life enzyme-containing hydrogel.

3. The method of claim 2, wherein the anti-swelling enzyme-containing hydrogel is prepared by: In step (1), the mass-volume ratio of the sulfobetaine monomer, methylene bisacrylamide, ammonium persulfate, carboxylated cellulose nanofiber, PBS buffer solution, and platinum nanoparticle solution is (0.3-0.5) g:(4-6) mg:(2-4) mg:(0.3-0.5) ml:(0.1-0.2) ml:(70-80) μl.

4. The use of the anti-swelling enzyme-containing hydrogel of claim 1 in the preparation of a diabetes detection device.

5. Use according to claim 4, characterized in that: The diabetes detection device is an electrochemical glucose enzyme sensor.

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