Preparation method and application of high-activity light-operated inactivated DNase-I nanoparticles

Abstract

The invention relates to a medical nano biological material, in particular to a preparation method and application of high-activity DNase-I nanoparticles capable of being inactivated in a light-operated mode. Strong electrostatic interaction occurs between a sulfonic negative charge group on a photosensitizer molecule and a positively charged region on the surface of DNase-I protein under the assistance of hydrogen bonds and hydrophobic interaction forces, so that the efficient photosensitizer and DNase-I co-loaded nanoparticles are formed, and the DNA degradation activity of DNase-I is remarkably enhanced. After nucleic acid pollutants in the environment are efficiently removed by the DNase-I nanoparticles, the loaded photosensitizer can be activated through illumination with specific wavelength such as an LED light source, and the active center of the DNase-I nanoparticles is irreversibly damaged by reactive oxygen species (ROS) generated by the photosensitizer, so that the activity of residual enzyme is thoroughly eliminated, and the problem of nuclease pollution is effectively solved.

Description

TECHNICAL FIELD

[0001] The application relates to a medical nanobiomaterial, in particular to a preparation method and application of high-activity light-controllable inactivated DNase-I nanoparticles. BACKGROUND

[0002] In molecular biology experiments, effectively preventing and completely eliminating deoxyribonucleic acid (DNA) pollution in the environment is a key link to ensure the accuracy of experimental results. The traditional removal method relying on disinfectants generally has the disadvantages of long action time and limited efficiency. In comparison, DNase-I, which has the ability to specifically hydrolyze DNA, has become an effective tool for eliminating such pollution. However, its high cost significantly limits its application, and therefore, improving the DNA degradation activity of DNase-I has become an important way to reduce operating costs.

[0003] The nanotechnology of biological enzyme molecules, that is, the combination of enzyme molecules with specific nanomaterials or the reconstruction of enzyme molecules into nanoscale structures, has been proven to significantly enhance the catalytic efficiency, stability and reusability of enzymes. This provides a promising strategy for solving the problem of improving the activity of DNase-I. However, this technology still faces core challenges when applied to DNase-I nanoparticles: how to maintain or even improve the enzyme activity to the greatest extent during nanofication, and how to achieve strict batch consistency during large-scale preparation.

[0004] Therefore, it is urgent to develop a new nanofication technology scheme that is relatively simple in preparation process and can effectively improve the activity of DNase-I nanoparticles. The core goal of this is to significantly enhance the activity of DNase-I, thereby reducing its use cost.

[0005] In addition, a commonly overlooked but crucial problem is that after DNase-I nanoparticles efficiently degrade residual DNA, they themselves remain as new protein contaminants. This residual nuclease seriously interferes with subsequent delicate operations involving nucleic acids, which may lead to false negative or false positive results, affecting the accuracy and repeatability of experiments. SUMMARY

[0006] To solve the problems raised in the background art, the application provides a preparation method and application of high-activity light-controllable inactivated DNase-I nanoparticles.

[0007] To achieve the above-mentioned purpose, the application provides the following technical scheme: a preparation method and application of high-activity light-controllable inactivated DNase-I nanoparticles, comprising the following steps:

[0008] S1, weighing DNase-I and configuring it into an aqueous solution;

[0009] S2, dissolving the photosensitizer in ultrapure water to prepare a solution;

[0010] S3, transferring the DNase-I aqueous solution to a flask, starting stirring on a magnetic stirrer, and adding the photosensitizer dropwise into the DNase-I aqueous solution;

[0011] S4, after uniform stirring, collecting the stirred reaction solution into a centrifuge tube, centrifuging and collecting the supernatant into another centrifuge tube, and retaining the reaction solution in the centrifuge tube for determination of the photosensitizer and DNase-I encapsulation rate;

[0012] S5, resuspending the precipitate with an appropriate amount of ultrapure water, and preparing high-activity light-controllable inactivated DNase-I nanoparticles by probe ultrasonic dispersion under an ultrasonic cell disruptor.

[0013] Preferably, the DNase-I is derived from a cow.

[0014] Preferably, the photosensitizer is a high-activity light-controllable inactivated DNase-I nanoparticle preparation method and application of one or more of a triphenylamine-based aggregation-induced emission molecule with a chemical structure containing a sulfonic acid group, and the number of sulfonic acid groups is between 1 and 3.

[0015] Preferably, the mass ratio of the DNase-I nanoparticles to the photosensitizer is 1:1-5:1.

[0016] Preferably, the prepared DNase-I nanoparticles have a particle size of 100-200 nm, and the encapsulation rates of DNase-I and the photosensitizer are both higher than 70%.

[0017] Preferably, the enzyme activator is used to remove DNA contamination.

[0018] Preferably, the DNA contamination sources include plasmid DNA, calf thymus DNA, spleen DNA, and PCR fragment DNA.

[0019] Preferably, the enzyme activator includes magnesium chloride and calcium chloride, and the concentrations of the two are 10 mM and 2 mM, respectively.

[0020] Preferably, after removing the DNA contamination, the photosensitizer in the DNase-I nanoparticles produces reactive oxygen species through LED irradiation, causing the DNase-I enzyme activity to be inactivated.

[0021] Compared with the prior art, the present application has the following beneficial effects:

[0022] The present application not only illustrates the key role of the photosensitizer in the formation of DNase-I nanoparticles, but also effectively solves the technical bottlenecks such as low loading rate of the photosensitizer and DNase-I and limited improvement of DNase-I enzyme activity that exist in the prior art.

[0023] The principle of action is that the sulfonic acid group negative charge on the photosensitizer molecule and the DNase-I nanoparticle protein surface with positive charged region strong electrostatic interaction, supplemented by hydrogen bond and hydrophobic force, so as to realize the efficient co-loading of photosensitizer and DNase-I, and significantly enhance the activity of DNase-I to degrade DNA.

[0024] After the DNase-I nanoparticles efficiently remove the nucleic acid pollutants in the environment, the loaded photosensitizer can be activated by LED light source and other specific wavelength light, and the active oxygen (ROS) generated by the photosensitizer can irreversibly damage the active center of the DNase-I nanoparticles, thereby completely eliminating the residual enzyme activity and effectively solving the problem of nuclease pollution. BRIEF DESCRIPTION OF DRAWINGS

[0025] The accompanying drawings are included to provide a further understanding of the application, and constitute a part of the specification, illustrate the application, and are used to explain the application, and do not constitute a limitation on the application. In the drawings:

[0026] Figure 1 The chemical structure of the sulfonated water-soluble AIE-type photosensitizer based on triphenylamine is shown;

[0027] Figure 2 The comparison chart of the removal effect of different concentrations of DNase-I / TRS1 nanoparticles on plasmid DNA is shown;

[0028] Figure 3 The comparison chart of the removal effect of different concentrations of DNase-I / TRS1 nanoparticles on calf thymus DNA is shown;

[0029] Figure 4 The comparison chart of the removal effect of different concentrations of DNase-I / TRS1 nanoparticles on spleen DNA is shown;

[0030] Figure 5 The comparison chart of the removal effect of different concentrations of DNase-I / TRS1 nanoparticles on PCR fragment DNA is shown;

[0031] Figure 6 The comparison chart of the removal effect of the same concentration of DNase-I / TRS2 and DNase-I nanoparticle / TRS3 nanoparticles on plasmid DNA and spleen DNA is shown;

[0032] Figure 7 The comparison chart of the removal effect of the same concentration of different DNase-I nanoparticles on plasmid DNA before and after light irradiation is shown; DETAILED DESCRIPTION

[0033] With reference to the accompanying drawings on the basis of the embodiments of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0034] Embodiment one:

[0035] A preparation method of high-activity light-controllable inactivated DNase-I nanoparticles and application of the high-activity light-controllable inactivated DNA scavenger are provided in the embodiment, which comprises the following components:

[0036] Table 1 Component table of the high-activity light-controllable inactivated DNA scavenger in embodiment one

[0037] Self-assembled DNase-I nanoparticles 0.01-50 μg / mL (as DNase-I) Magnesium chloride 10 mM Calcium chloride 2 mM

[0038] Specifically, the self-assembled DNase-I nanoparticles are prepared by the following preparation method:

[0039] S1, DNase-I is weighed and configured into an aqueous solution with a concentration of 2 mg / mL;

[0040] S2, the mother liquor of TRS1 is diluted to 1 mg / mL;

[0041] S3, the DNase-I aqueous solution (1000 μL) is transferred to a test tube, and stirring is started on a magnetic stirrer;

[0042] S4, the TRS1 aqueous solution is added dropwise into the test tube by using a 1 mL syringe;

[0043] S5, after stirring for 30 min, the stirred reaction solution is collected into a 5 mL centrifuge tube, centrifuged (10000 g, 5 min), and the supernatant is collected into another centrifuge tube, which is reserved for determination of the TRS1 and DNase-I encapsulation rate;

[0044] S6, the precipitate is resuspended with an appropriate amount of water, and the prepared nanoparticles are dispersed under probe ultrasonication (100 W, 1 min) in an ultrasonic cell disruptor;

[0045] S7, it is determined that the particle size is 196±0.82 nm, the PDI (polydispersity index) is 0.134±0.008, the DNase-I encapsulation rate is 99.3±0.003%, and the photosensitizer TRS1 encapsulation rate is 99.7±0.003%.

[0046] Embodiment two:

[0047] The present embodiment provides a preparation method of high-activity light-controllable inactivated DNase-I nanoparticles and application of high-activity light-controllable inactivated DNA scavenger, which comprises the following components:

[0048] Table 2 Component table of high-activity light-controllable inactivated DNA scavenger in Example 2

[0049] Self-assembled DNase-I nanoparticles 0.01-50 μg / mL (as DNase-I) Magnesium chloride 10 mM Calcium chloride 2 mM

[0050] The preparation method of the self-assembled DNase-I nanoparticles in Example 2 is consistent with that in Example 1, except that TRS1 is replaced by TRS2, wherein TRS1 and TRS2 are photosensitizers from different sources.

[0051] It is determined that the particle size is 159±0.58 nm, the PDI (polydispersity index) is 0.118±0.006, the DNase-I encapsulation rate is 98.1±0.009%, and the photosensitizer TRS2 encapsulation rate is 98.1±0.005%.

[0052] Example 3:

[0053] The present embodiment provides a preparation method of high-activity light-controllable inactivated DNase-I nanoparticles and application of high-activity light-controllable inactivated DNA scavenger, which comprises the following components:

[0054] Table 3 Component table of high-activity light-controllable inactivated DNA scavenger in Example 3

[0055] Self-assembled DNase-I nanoparticles 0.01-50 μg / mL (as DNase-I) Magnesium chloride 10 mM Calcium chloride 2 mM

[0056] The preparation method of the self-assembled DNase-I nanoparticles in Example 2 is consistent with that in Example 1, except that TRS1 is replaced by TRS3, wherein TRS1 and TRS3 are photosensitizers from different sources.

[0057] It is determined that the particle size is 145±1.05 nm, the PDI (polydispersity index) is 0.123±0.008, the DNase-I encapsulation rate is 72.2±0.121%, and the photosensitizer TRS3 encapsulation rate is 87.3±0.041%.

[0058] Comparative Example 1:

[0059] The present embodiment provides a common DNase-I scavenger, which comprises the following components:

[0060] Table 4 Component table of common DNase-I scavenger in Comparative Example 1

[0061] DNase-I solution 0.01-50 μg / mL Magnesium chloride 10 mM Calcium chloride 2 mM

[0062] The preparation method of the DNase-I solution is:

[0063] DNase-I was weighed and dissolved in water to a concentration of 2 mg / mL;

[0064] The DNA scavengers in Examples 1-3 were verified for performance by the following experiment:

[0065] Experimental Example:

[0066] Example 1 and Comparative Example 1 were verified for the ability to remove plasmid DNA:

[0067] Ten identical 0.2 mL EP tubes were prepared, labeled a-j.

[0068] Plasmid DNA (200 μg / mL, 2 μL) was added to tubes b-j, and Comparative Example 1 (8 μL) was added to tubes c-f, containing DNase-I at concentrations of 10, 2, 1, and 0.5 μg / mL, respectively. Example 1 (8 μL) was added to tubes g-j, containing DNase-I at concentrations of 10, 2, 1, and 0.5 μg / mL, respectively. Tube b was supplemented with TAE (Tris-acetate-EDTA) to a final concentration of 10 mM MgCl2 and 2 mM CaCl2 as a control.

[0069] The reaction was then carried out at 37°C for 20 min.

[0070] The samples in the 10 EP tubes were detected by 0.7% agarose gel electrophoresis, and the images were observed under a gel imager, as shown in Figure 2 .

[0071] Figure 2 The results show the effects of different concentrations of DNase-I / TRS1 nanoparticles of Example 1 on the removal of plasmid DNA.

[0072] Lane a: DNA Marker (250-10,000 bp);

[0073] Lane b: Plasmid DNA;

[0074] Lanes c-f: Plasmid DNA + Comparative Example 1;

[0075] Lanes g-j: Plasmid DNA + Example 1.

[0076] As can be seen from Figure 2 , nucleic acids still exist in lanes b-f, while no bright bands are observed in lanes g-j containing DNase-I / TRS1 nanoparticles, indicating that the nucleic acids in lanes g-j are completely digested, thus indicating that the DNase-I / TRS1 nanoparticle has better enzymatic activity than free DNase-I nanoparticles and can degrade plasmid DNA.

[0077] Example 1 and Comparative Example 1 verify the removal ability of calf thymus DNA:

[0078] Prepare 11 identical 0.2 mL EP tubes, respectively marked as a-k tubes;

[0079] Add calf thymus DNA (400 μg / mL, 2 μL) into b-k tubes, add Comparative Example 1 8 μL into b-e tubes, which contains DNase-I with concentrations of 50, 10, 5, 2.5 μg / mL, respectively, add Example 1 8 μL into f-i tubes, which contains DNase-I nanoparticles with concentrations of 50, 10, 5, 2.5 μg / mL, respectively, and add TAE to j, k tubes to make up to 10 μL, so that the final concentrations of magnesium chloride and calcium chloride are 10 mM and 2 mM, respectively, as controls;

[0080] Then react at 37°C for 20 min.

[0081] Detect the samples in the 11 EP tubes by 0.7% agarose gel electrophoresis, and observe the images under a gel imager, and the results are shown in Figure 3 .

[0082] Figure 3 The figure shows the removal effects of calf thymus DNA by DNase-I / TRS1 nanoparticles with different concentrations according to Example 1.

[0083] Lane a: DNA Marker (250-10000 bp);

[0084] Lanes b-e: calf thymus DNA + Comparative Example 1;

[0085] Lanes f-i: calf thymus DNA + Example 1;

[0086] Lanes j-k: calf thymus DNA.

[0087] As can be seen from Figure 3 , nucleic acids still exist in lanes b-e, while no bright bands are observed in lanes f-i which contain DNase-I / TRS1 nanoparticles, indicating that the nucleic acids in lanes f-i are completely digested, thus indicating that the DNase-I / TRS1 nanoparticle has better enzymatic activity than free DNase-I nanoparticles and can degrade calf thymus DNA.

[0088] Example 1 and Comparative Example 1 verify the removal ability of calf thymus DNA:

[0089] Prepare 16 identical 0.2 mL EP tubes, respectively marked as 1-16 tubes;

[0090] Before adding spleen DNA and PCR fragment (4 μL) into tube 2-8 and 10-16, respectively, adding 1 μL of Example 1 into tube 2-4 and 10-12, containing DNase-I at the concentration of 1, 0.1, 0.01 μg / mL, respectively, adding 1 μL of Example 1 into tube 5-7 and 13-15, containing DNase-I nanoparticles at the concentration of 1, 0.1, 0.01 μg / mL, respectively, wherein tube 8 and 16 are supplemented with TAE to 5 μL, so that the final concentration of magnesium chloride and calcium chloride is 10 mM and 2 mM, respectively, as a control;

[0091] Then, reacting at 37°C for 20 min.

[0092] The samples in the 16 EP tubes were detected by 0.7% agarose gel electrophoresis, and the images were observed under a gel imager, and the results are shown in Figure 4 and Figure 5 .

[0093] Figure 4 The figure shows the comparison of the removal effect of different concentrations of DNase-I / TRS1 nanoparticles according to Example 1 of the present application on spleen DNA.

[0094] Lane a: DNA Marker (250-10000 bp);

[0095] Lanes b-d: Spleen DNA + Comparative Example 1;

[0096] Lanes e-g: Spleen DNA + Example 1;

[0097] Lane h: Spleen DNA.

[0098] As can be seen from Figure 4 , in lane b, nucleic acids still exist, while in lane e, no bright band is observed after adding DNase-I / TRS1 nanoparticles, indicating that the nucleic acids in lane e are completely digested, thereby indicating that the DNase-I / TRS1 nanoparticle has better enzyme activity than free DNase-I nanoparticles and can degrade spleen DNA.

[0099] Figure 5 The figure shows the comparison of the removal effect of different concentrations of DNase-I / TRS1 nanoparticles according to Example 1 of the present application and DNase-I according to the comparative example on PCR fragments.

[0100] Lane a: DNA Marker (250-10000 bp);

[0101] Lanes b-d: PCR fragment + Comparative Example 1;

[0102] Lanes e-g: PCR fragment + Example 1;

[0103] Lane h: PCR fragment.

[0104] As can be seen from Figure 5 , the nucleic acid still exists in lane b, while no bright band is observed in lane e with the addition of DNase-I / TRS1 nanoparticles, indicating that the nucleic acid digestion in lane e is complete, thus indicating that the DNase-I / TRS1 nanoparticle has better enzyme activity than the free DNase-I nanoparticle and can degrade the PCR fragment.

[0105] Example 2, Example 3 and Comparative Example 1 for verifying the removal ability of spleen DNA and plasmid DNA:

[0106] Nine identical 0.2 mL EP tubes were prepared, and were respectively marked as a-i tubes;

[0107] Spleen DNA (400 μg / mL, 4 μL) was added to the b-e tubes, and Comparative Example 1, Example 2 and Example 3 were respectively added to the c-e tubes, each 1 μL, containing DNase-I at a concentration of 0.5 μg / mL. Plasmid DNA (439 μg / mL, 4 μL) was added to the f-i tubes, and Comparative Example 1, Example 2 and Example 3 were respectively added to the g-i tubes, each 1 μL, containing DNase-I at a concentration of 0.5 μg / mL. The b and f tubes were supplemented with TAE to 5 μL, so that the final concentrations of magnesium chloride and calcium chloride were 10 mM and 2 mM, respectively, as controls;

[0108] Then, 37°C reaction was carried out for 20 min.

[0109] The samples in the nine EP tubes were detected by 0.7% agarose gel electrophoresis, and the images were observed under a gel imaging instrument, and the results are shown in Figure 6 .

[0110] Figure 6 The removal effects of the DNase-I / TRS2 nanoparticles and the DNase-I / TRS3 nanoparticles according to Example 2 and Example 3 of the present application on spleen DNA and plasmid DNA are shown.

[0111] Lane a: DNA Marker (250-10000 bp);

[0112] Lane b: Spleen DNA; Lane c: Spleen DNA + Comparative Example 1;

[0113] Lane d: Spleen DNA + Example 2;

[0114] Lane e: Spleen DNA + Example 3;

[0115] Lane f: Plasmid DNA;

[0116] Lane g: Plasmid DNA + Comparative Example 1;

[0117] Lane h: Plasmid DNA + Example 2;

[0118] Lane i: Plasmid DNA + Example 3.

[0119] from Figure 6 As can be seen, nucleic acids were still present in lanes bc and fg, while no bright bands were observed in lanes de and hi, which contained DNase-I / TRS2 and DNase-I / TRS3 nanoparticles, respectively. This indicates that the nucleic acids in lanes de and hi were completely digested, thus demonstrating that the enzyme activity of the two nanoparticles is superior to that of free DNase-I nanoparticles and that they can degrade plasmid DNA and PCR fragments.

[0120] Verification of photo-controlled quenching activity:

[0121] Prepare 10 identical 0.2mL EP tubes, and label them tube number aj;

[0122] First, add plasmid DNA (312 μg / mL, 4 μL) to tube bj. Then, add 1 μL each of Comparative Example 1, Example 1, Example 2, and Example 3 to tube cf, each containing DNase-I at a concentration of 20 μg / mL. Finally, add Comparative Example 1, Example 1, Example 2, and Example 3 (all treated with light for 5 min at 0.33 W / cm²) to tube gj. 2 Each tube contains 1 μL of DNase-I at a concentration of 20 μg / mL. Tubes b and f are supplemented with TAE to a final volume of 5 μL, so that the final concentrations of magnesium chloride and calcium chloride are 10 mM and 2 mM, respectively, as controls.

[0123] Then react at 37°C for 20 minutes.

[0124] The samples in 10 EP tubes were examined by 0.7% agarose gel electrophoresis, and the images were observed under a gel imaging system. The results are as follows. Figure 7 As shown.

[0125] Figure 7 The photo-controlled inactivation performance of the present invention is demonstrated.

[0126] Lane a: DNA Marker (250–10000 bp);

[0127] Lane b: Plasmid DNA;

[0128] Lane c: Plasmid DNA + Comparative Example 1;

[0129] Lane d: Plasmid DNA + Example 1;

[0130] Lane e: Plasmid DNA + Example 2;

[0131] Lane f: plasmid DNA + Example Three;

[0132] Lane g: plasmid DNA + Comparative Example One (after light);

[0133] Lane h: plasmid DNA + Example One (after light);

[0134] Lane i: plasmid DNA + Example Two (after light);

[0135] Lane j: plasmid DNA + Example Three (after light).

[0136] As can be seen from Figure 7 lanes b-c and g-j, the nucleic acid still exists, while no bright band is observed in lanes d-f of DNase-I / TRS1 nanoparticle, DNase-I / TRS2 nanoparticle and DNase-I / TRS3 nanoparticle without light, indicating that the nucleic acid in lanes d-f is completely digested, thus indicating that the three kinds of nanoparticle enzymes cannot degrade DNA after light, while the common DNase-I has poor degradation effect on plasmid DNA, which does not degrade DNA before and after light.

[0137] Finally, it should be noted that the above only describes the preferred embodiments of the present application and is not intended to limit the present application. Although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent replacements to some technical features. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A method for preparing highly active, photo-controlled inactivation-prone DNase-I nanoparticles, characterized in that, Includes the following steps: S1. Weigh out DNase-I and prepare it into an aqueous solution; S2. Dissolve the photosensitizer in ultrapure water to prepare a solution; S3. Transfer the DNase-I aqueous solution to a vial, start stirring on a magnetic stirrer, and add the photosensitizer dropwise to the DNase-I aqueous solution; S4. After stirring evenly, collect the stirred reaction solution into a centrifuge tube, centrifuge, and collect the supernatant into another centrifuge tube. The reaction solution remaining in the centrifuge tube is used to determine the photosensitizer and DNase-I encapsulation rate. S5. The precipitate is resuspended in an appropriate amount of ultrapure water and then dispersed by ultrasonication under the probe of an ultrasonic cell disruptor to obtain highly active DNase-I nanoparticles that can be photo-controlled to be inactivated.

2. The method for preparing highly active, photo-controlled inactivation-prone DNase-I nanoparticles according to claim 1, characterized in that: The DNase-I is derived from bovine tannins.

3. The method for preparing highly active, photo-controlled inactivation-prone DNase-I nanoparticles according to claim 1, characterized in that: The photosensitizer is one or more of triphenylamine-based aggregation-induced emission molecules with a chemical structure containing sulfonic acid groups, and the number of sulfonic acid groups is between 1 and 3.

4. The method for preparing highly active, photo-controlled inactivation-prone DNase-I nanoparticles according to claim 1, characterized in that: The mass ratio of DNase-I nanoparticles to photosensitizer is 1:1 to 5:

1.

5. The method for preparing highly active, photo-controlled inactivation-prone DNase-I nanoparticles according to claims 1-4, characterized in that: The prepared DNase-I nanoparticles have a particle size of 100-200 nm; the encapsulation efficiency of both DNase-I and photosensitizer is higher than 70%.

6. An application of highly active, photo-inactivated DNase-I nanoparticles prepared according to any one of claims 1-4, characterized in that: It works in conjunction with enzyme activators to remove DNA contamination.

7. The application of the DNase-I nanoparticles according to claim 6 in the removal of DNA contamination, characterized in that: DNA contamination sources include plasmid DNA, calf thymus DNA, spleen DNA, and PCR fragment DNA.

8. The application of the DNase-I nanoparticles according to claim 6 in the removal of DNA contamination, characterized in that: The enzyme activator includes magnesium chloride and calcium chloride, with concentrations of 10 mM and 2 mM, respectively.

9. The application of the DNase-I nanoparticles according to claim 6 in the removal of DNA contamination, characterized in that: After DNase-I nanoparticles remove DNA contamination, LED light irradiation causes the photosensitizer in the DNase-I nanoparticles to generate reactive oxygen species, thereby inactivating the DNase-I enzyme.

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