Immunoglobulin g nanoparticle and industrial preparation method and application thereof

Abstract

The application relates to the technical field of immunoglobulin G nanoparticle preparation, in particular to an immunoglobulin G nanoparticle and a preparation method and application thereof; the application discloses an immunoglobulin G nanoparticle, wherein the particle size of the immunoglobulin G nanoparticle is 10 nm-200 nm; the application discloses an immunoglobulin G nanoparticle and a preparation method and application thereof, the immunoglobulin G nanoparticle prepared by the method has better thermal stability, and the immunoglobulin G nanoparticle prepared still maintains its effectiveness under high temperature; compared with traditional immunoglobulin G, the immunoglobulin G nanoparticle prepared by the method of the application effectively prolongs the half-life period and does not need to be frequently taken in; the immunoglobulin G nanoparticle prepared by the method of the application has higher biological activity.

Description

Technical Field

[0001] This application relates to the technical field of immunoglobulin G nanoparticle preparation, and in particular to an immunoglobulin G nanoparticle and its industrial preparation method and application. Background Technology

[0002] Bovine colostrum contains a wealth of nutrients and bioactive components, including immunoglobulins, growth factors, hormones, enzymes, lactoferrin, cytokines, leukocytes, nucleosides, and nucleotides. Immunoglobulin G (IgG) is the most abundant antibody in bovine colostrum, accounting for 80-85% of the total immunoglobulins. It comprises two subclasses: IgG1 (the most abundant immunoglobulin isotype) and IgG2. Before the invention of artificial antibodies, bovine colostrum was considered the key to unlocking the mechanisms of preventing microbial infections. Immunoglobulins are well-known for their beneficial role in immunogenic responses in human health. When a host encounters a foreign substance (antigen), antibodies bind to, recognize, and destroy bacteria, toxins, viruses, and other antigens. When the antigen re-enters the body, it stimulates the production of the same antibodies to clear the infection.

[0003] Temperature, pH, enzymatic degradation, and interactions with other components can affect the activity of immunoglobulins. To address these issues, many nanoparticles synthesized from non-biological composite materials are beginning to be applied in biomedicine, such as drug delivery, enzyme immobilization, diagnostic imaging, cancer therapy, and targeted gene delivery. Nanoparticles offer better stability, targeted delivery, and enhanced bioactivity, and can significantly impact human health.

[0004] Therefore, how to prepare immunoglobulin G into nanoparticles with thermal stability, biological activity, and a longer half-life has become an urgent problem to be solved. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this application provides immunoglobulin G nanoparticles, their industrial preparation method, and their applications.

[0006] This application aims to solve the technical problems of immunoglobulin's instability at high temperatures, short half-life, and loss of bioactivity. By extracting IgG nanoparticles from bovine colostrum, its effectiveness can be maintained, its lifespan extended, and its bioactivity enhanced. This allows it to be applied in various biomedical fields, including beverages, treatments, and other product manufacturing processes.

[0007] In a first aspect, this application provides an immunoglobulin G nanoparticle, which adopts the following technical solution: An immunoglobulin G nanoparticle, wherein the particle size of the immunoglobulin G nanoparticle is 10 nm-200 nm.

[0008] Conventional immunoglobulin G (i.e., conventional IgG without nanoprocessing) has a particle size >500nm, and immunoglobulin G is prone to aggregate with proteins or itself to form polymerized IgG.

[0009] The immunoglobulin G nanoparticles prepared in this application have a particle size of 10nm-200nm. The nanoscale immunoglobulin IgG prepared in this application does not aggregate with proteins or itself, so the prepared immunoglobulin G nanoparticles have more exposed surface sites, which can more effectively bind pathogens and promote their inactivation.

[0010] Preferably, the immunoglobulin G nanoparticles are extracted from bovine colostrum.

[0011] Secondly, this application provides an industrial preparation method for immunoglobulin G nanoparticles, employing the following technical solution: An industrial preparation method for immunoglobulin G nanoparticles, comprising the following steps: (1) The bovine colostrum was centrifuged to remove fat, and the result was defatted bovine colostrum; (2) Heat the skimmed bovine colostrum to 40-50℃, adjust the pH of the skimmed bovine colostrum to 4.4-4.6, filter, remove casein, and obtain whey; (3) Adjust the pH of the whey to 3-3.8, filter, and obtain purified whey; (4) The purified whey was concentrated by ultrafiltration to obtain nanoparticles; (5) The nanoparticles were dried to obtain immunoglobulin G nanoparticles.

[0012] By adopting the above technical solution, the immunoglobulin G nanoparticles prepared in this application through centrifugation defatting, removal of casein, whey purification, ultrafiltration concentration, and drying are characterized by good dispersibility, thermal stability, hydrophobicity, and nonpolarity.

[0013] In this application, the pH of skimmed bovine colostrum is first adjusted using an acid solution to precipitate casein. The casein is then removed by filtration to obtain whey. By adjusting the pH of the whey, it is further purified to further remove protein precipitates and obtain purer whey.

[0014] Preferably, in step (1), the temperature of the bovine colostrum is controlled at 4-18°C before centrifugation and defatting.

[0015] By adopting the above technical solution, the temperature of bovine colostrum is controlled at 4-18℃ before centrifugation and defatting, which can ensure the quality and biological activity of bovine colostrum and effectively prevent bacteria in the colostrum from affecting its quality.

[0016] Preferably, during the centrifugation defatting in step (1), the centrifugation speed is 3000-6000 rpm, and the temperature of the bovine colostrum is controlled to be ≤25℃ during the centrifugation process.

[0017] By adopting the above technical solution, the temperature of bovine colostrum is controlled during centrifugation. Lower temperatures help separate fats and effectively prevent fat liquefaction caused by increased temperature, which would affect separation efficiency.

[0018] Preferably, in step (2), the solution used to adjust the pH of the skimmed bovine colostrum is an acidic solution.

[0019] Preferably, the acid solution used is hydrochloric acid solution, citric acid solution, vitamin C solution or acetic acid solution, etc.

[0020] Preferably, in step (2), the filtration step is as follows: the pH-adjusted skimmed bovine colostrum is filtered through a 60-200 mesh sieve.

[0021] Preferably, in step (3), the solution used to adjust the pH of the whey is an acidic solution.

[0022] Preferably, the acid solution used is hydrochloric acid solution, citric acid solution, vitamin C solution or acetic acid solution, etc.

[0023] Preferably, in step (3), the filtration step is as follows: the whey after pH adjustment is filtered through a sieve with a mesh size of ≥200.

[0024] Preferably, in step (4), during ultrafiltration concentration, an ultrafiltration device with a membrane molecular weight cutoff of 10 kDa is used for ultrafiltration concentration.

[0025] Preferably, the immunoglobulin G nanoparticles obtained after drying in step (5) have a particle size of 10nm-200nm.

[0026] Preferably, the drying method in step (5) includes various drying methods such as freeze drying and spray drying.

[0027] Thirdly, this application provides an application of immunoglobulin G nanoparticles or immunoglobulin G nanoparticles prepared by the above method in the preparation of food, health products or pharmaceuticals, using the following technical solution: Application of an immunoglobulin G nanoparticle or immunoglobulin G nanoparticles prepared by the above method in the preparation of food, health products or pharmaceuticals.

[0028] The immunoglobulin G nanoparticles prepared using the method described in this application have structural stability and high antibody activity, and can be widely used in food, health products, liquid beverages or other products.

[0029] In summary, this application includes at least one of the following beneficial technical effects: 1. This application discloses an immunoglobulin G nanoparticle and its industrial preparation method and application. The prepared immunoglobulin G nanoparticle has better thermal stability and retains its effectiveness at high temperatures. 2. The immunoglobulin G nanoparticles prepared using the method of this application have an effectively extended half-life compared with traditional immunoglobulin G; 3. The immunoglobulin G nanoparticles prepared using the method of this application have higher biological activity. Attached Figure Description

[0030] Figure 1 Transmission electron microscopy (TEM) image of immunoglobulin G nanoparticles extracted using the method of Example 1; Figure 2 The images show scanning electron microscopy (SEM) images of immunoglobulin G nanoparticles extracted using the method in Example 1 and conventional IgG without nanoprocessing; wherein, Figure 2 The left image is a SEM image of IgG nanoparticles. Figure 2 The image on the right is a SEM image of conventional IgG without nano-processing; Figure 3 Size distribution map of immunoglobulin G nanoparticles extracted using the method of Example 1, as displayed by dynamic light scattering; Figure 4 The thermostability graphs show the immunoglobulin G nanoparticles extracted using the method in Example 1 and conventional IgG without nano-sizing treatment; wherein, Figure 4 (a) shows the thermal stability of IgG nanoparticles; Figure 4 (b) Thermal stability diagram of conventional IgG without nano-processing; Figure 5 The images show denaturation patterns of immunoglobulin G nanoparticles extracted using the method in Example 1 and conventional IgG without nano-processing; wherein, Figure 5 (a) is a denaturation diagram of IgG nanoparticles; Figure 5 (b) Denaturation diagram of conventional IgG without nano-processing; Figure 6 An aggregate diagram of immunoglobulin G nanoparticles extracted using the method of Example 1 and conventional IgG without nanoprocessing; Figure 7 The denaturation detection results are for immunoglobulin G nanoparticles extracted using the method in Example 1 and conventional IgG without nano-processing. Figure 8The following are the Fourier Transform Infrared (FTIR) spectra of immunoglobulin G nanoparticles extracted using the method of Example 1 and conventional IgG without nano-processing. Figure 9 X-ray diffraction (X-RD) patterns of immunoglobulin G nanoparticles extracted using the method of Example 1 and conventional IgG without nano-processing. Figure 10 Thermogravimetric analysis (TGA) images of immunoglobulin G nanoparticles extracted using the method of Example 1 and conventional IgG without nano-processing are shown. Detailed Implementation

[0031] The technical solutions of this application are further illustrated by specific embodiments below. These specific embodiments do not represent a limitation on the scope of protection of this application. Any non-essential modifications and adjustments made by others based on the concept of this application still fall within the scope of protection of this application.

[0032] All raw materials involved in this application are commercially available products, among which, Conventional IgG without nanoprocessing, i.e. reagent-grade purified human immunoglobulin G, Sigma-Aldrich (USA).

[0033] The present application will be further described in detail below with reference to embodiments and comparative examples.

[0034] An industrial preparation method for immunoglobulin G nanoparticles, comprising the following steps: Step 1: Collect bovine colostrum: Milk the cows and collect the bovine colostrum, controlling its temperature to 4-18℃; Step 2, centrifugation and defatting: The bovine colostrum is transported to a centrifuge for defatting to obtain defatted bovine colostrum; The centrifugal defatting machine has a current setting of 7.5-8.5A and a speed of 3000-6000rpm; the temperature of bovine colostrum is controlled to be ≤25℃ during the centrifugal defatting process.

[0035] Step 3: Remove casein: Heat the skimmed bovine colostrum to 40-50℃ and adjust the pH of the skimmed bovine colostrum to 4.4-4.6 using an acid solution (such as hydrochloric acid solution, citric acid solution, vitamin C solution, or acetic acid solution). At this point, the casein reaches its isoelectric point and begins to coagulate, causing the casein to precipitate. Filter the precipitate through a 60-200 mesh sieve to remove the casein and obtain whey. The step of filtering through a 60-200 mesh sieve in this process can be replaced by centrifugation.

[0036] Step 4: Whey purification: The obtained whey is purified to obtain purified whey; Specifically, the pH of the whey is adjusted to 3-3.8 using an acid solution (such as hydrochloric acid solution, citric acid solution, vitamin C solution, or acetic acid solution), and then filtered through a ≥200 mesh sieve to finally obtain purified whey. The step of filtering through a 200-mesh screen in this process can be replaced by centrifugation.

[0037] Step 5, Ultrafiltration Concentration: The purified whey is concentrated by ultrafiltration to remove lactose, small molecules and regularly dispersed immunoglobulin IgG, and to convert immunoglobulin IgG into nanoparticles. Among them, ultrafiltration equipment is used for ultrafiltration concentration, and the membrane used in the ultrafiltration equipment has a molecular weight cutoff of 10 kDa.

[0038] Step 6: Drying: The obtained nanoparticles are dried to obtain immunoglobulin G nanoparticles; the particle size of the immunoglobulin G nanoparticles is 10nm-200nm.

[0039] Example 1: Immunoglobulin G nanoparticles extracted from bovine colostrum An industrial-scale preparation method for immunoglobulin G nanoparticles extracted from bovine colostrum, comprising the following steps: (1) Collecting bovine colostrum: Milk the cows and collect the colostrum, then immediately refrigerate at 4°C. (2) Centrifugal defatting: Bovine colostrum is transported to a centrifugal defatting machine for defatting to obtain defatted bovine colostrum. The current setting of the centrifugal defatting machine is 8.2A, the speed is 5000rpm, and the initial temperature of the milk during centrifugation is controlled to be ≤25℃; (3) Removal of casein: The temperature of the skimmed bovine colostrum was raised to 45°C, and the pH of the skimmed bovine colostrum was adjusted to 4.6 using 0.1M citric acid solution to precipitate casein. The casein was then removed by filtering through a 120-mesh sieve to obtain whey. (4) Whey purification: The pH of the whey was adjusted to 3.8 using 0.1M citric acid solution, and then filtered through a 200-mesh sieve to obtain purified whey.

[0040] (5) Ultrafiltration Concentration: Purified water was added to the ultrafiltration equipment for ultrafiltration. The membrane used in the ultrafiltration equipment had a molecular weight cutoff of 10 kDa. The purified whey obtained in step (4) was added to 2 times the volume of ultrapure water and ultrafiltered to the original volume through the 10 kDa ultrafiltration membrane. The retentate was recovered, and 2 times the original volume of ultrapure water was added. Ultrafiltration was then performed again. This process was repeated 3 times or more until the absorbance of the permeate measured at 280 nm by a UV spectrophotometer was less than 0.05. Then the permeate was concentrated.

[0041] (6) Drying: The obtained ultrafiltration concentrate was freeze-dried for 48 hours to obtain immunoglobulin G nanoparticles (i.e., IgG nanoparticles).

[0042] Example 2: The difference from Example 1 is that the pH of the whey is adjusted to 3.5 in step (4).

[0043] Example 3: The difference from Example 1 is that the pH of the whey is adjusted to 3 in step (4).

[0044] Comparative Example 1: The difference from Example 1 is that the pH of the whey is adjusted to 3.9 in step (4).

[0045] Comparative Example 2: The difference from Example 1 is that the pH of the whey is adjusted to 2.9 in step (4).

[0046] Immunoglobulin G nanoparticles were extracted from 100 kg of bovine milk using the above method. The immunoglobulin G nanoparticles obtained in Examples 1-3, Comparative Example 1, and Comparative Example 2 were measured using transmission electron microscopy (TEM). The results are shown in Table 1.

[0047] Table 1 Test Results Based on the test results in Table 1, it can be seen that the size distribution of the immunoglobulin G nanoparticles prepared in Examples 1-3 is between 10 nm and 200 nm.

[0048] Based on Example 1 and Comparative Example 1, it can be seen that although the yield of Comparative Example 1 is greater than that of immunoglobulin G nanoparticles, the particle size of Comparative Example 1 is greater than that of Example 1. This is mainly because the pH increases, which causes the prepared immunoglobulin G nanoparticles to contain a certain amount of impurities, thus making the yield of Comparative Example 1 higher than that of Example 1. Therefore, the detection results of Example 1 are better.

[0049] Based on Example 1 and Comparative Example 2, it can be seen that the yield of Comparative Example 2 is less than that of Comparative Example 1, so the test results of Example 1 are better.

[0050] Therefore, according to Table 1, adjusting the pH of whey to 3-3.8 during whey purification yields the best immunoglobulin G nanoparticles, and the immunoglobulin G nanoparticles prepared in Example 1 have the highest yield.

[0051] Performance testing: The immunoglobulin G nanoparticles prepared in Example 1 were measured.

[0052] 1. Transmission electron microscopy (TEM) measurements: The immunoglobulin G nanoparticles obtained in Example 1 were measured using transmission electron microscopy (TEM). The results showed that the size distribution of the immunoglobulin G nanoparticles ranged from 10 nm to 200 nm, with each immunoglobulin G nanoparticle measuring 10 nm. Most immunoglobulin G nanoparticles existed in clusters, with each cluster containing four to ten nanoparticles, each no larger than 200 nm. For detailed results, see [link to detailed results]. Figure 1 .

[0053] 2. Scanning electron microscopy (SEM) measurement The morphology of the immunoglobulin G nanoparticles prepared in Example 1 was characterized using scanning electron microscopy (SEM). Figure 2 The morphology of immunoglobulin G nanoparticles is compared with that of conventional IgG without nanoprocessing.

[0054] Figure 2 Left image (IgG nanoparticles): This image is a SEM image of IgG nanoparticles, magnified 350,000 times. The microstructure of the nanoparticles can be clearly observed in the image. The particles exhibit regular small spherical shapes and are tightly packed, indicating that these IgG particles have been successfully nanoscaled.

[0055] Figure 2 Right image (conventional IgG without nanoparticle processing): This image is a SEM image of conventional IgG without nanoparticle processing, magnified 400 times. The image shows a large, irregular sheet-like structure, indicating that this IgG sample has a large particle size and a relatively smooth surface, significantly different from the fine structure of nanoparticles.

[0056] pass Figure 2 It can be seen that the IgG nanoparticles prepared by the method of this application are more uniform and fine, while conventional IgG without nano-sizing treatment exhibits a larger block or sheet-like morphology, further proving the effectiveness of the nano-sizing process.

[0057] 3. Dynamic light scattering (DLS) measurement The immunoglobulin G nanoparticles obtained in Example 1 were further measured using a dynamic light scattering particle size analyzer (DLS). The DLS results confirmed that the size of the immunoglobulin G nanoparticles ranged from 10 nm to 200 nm. Figure 3 As shown, the DLS measurement results not only provide the average particle size, but also show the distribution range and uniformity of particle size, proving that the immunoglobulin G nanoparticles maintained good dispersibility and stability during the preparation process.

[0058] 4. Thermal stability test (1) Wavelength and absorbance detection The wavelength and absorbance of the immunoglobulin G nanoparticles prepared in Example 1 and conventional IgG without nano-sizing were detected using a UV-Vis spectrometer after heating at 85°C for 0, 5 min, 10 min, 20 min, 40 min, 60 min, and 120 min, respectively. The detection results are as follows: Figure 4 As shown.

[0059] according to Figure 4 It can be seen that the absorbance of immunoglobulin G nanoparticles varies little at different time points, which indicates that the sample has high stability in the ultraviolet-visible region (200-400nm) after nano-sizing.

[0060] Comparing the absorbance curves at different time points, the absorbance of immunoglobulin G nanoparticles showed little change during heat treatment (85℃), while the absorbance of conventional IgG without nano-sizing treatment showed a significant decreasing trend with increasing time. This indicates that nano-sizing treatment can improve the stability of IgG at high temperatures and slow down its denaturation or degradation.

[0061] The absorbance of unprocessed conventional IgG showed significantly greater changes at different time points (especially 5 minutes, 10 minutes and beyond), indicating that its structural stability was poor and it was more prone to denaturation or polymerization under high temperature conditions. Furthermore, the absorbance of unprocessed conventional IgG gradually decreased over time, possibly due to structural denaturation caused by high temperature, and the possibility that some IgG molecules may have undergone cross-linking reactions with other molecules, thus affecting absorbance. In contrast, nano-processed IgG exhibited higher resistance to denaturation under the same conditions, indicating that it had better structural integrity and a lower tendency to polymerization during heat treatment.

[0062] As heating time increased, the absorbance of conventional IgG without nano-processing decreased rapidly in the early stage (within 10 minutes), and then the change in absorbance tended to be gradual. This indicates that IgG underwent a relatively violent denaturation reaction in the initial stage, and then gradually reached a relatively stable state; while the absorbance of nano-IgG particles showed smaller changes at various time points, indicating that it maintained a relatively stable state throughout the heating process.

[0063] The immunoglobulin G nanoparticles prepared in Example 1 and conventional IgG without nano-treatment were heated at 85°C for 0, 5 min, 10 min, 20 min, 40 min, 60 min, and 120 min, respectively. The denaturation of the immunoglobulin G nanoparticles and conventional IgG without nano-treatment was observed. The denaturation results of the immunoglobulin G nanoparticles and conventional IgG without nano-treatment are shown below. Figure 5 As shown.

[0064] according to Figure 5 It is known that conventional IgG without nano-processing denatures and precipitates when heated at 85°C for 5 minutes, and all precipitates when heated for 40 minutes. The immunoglobulin G nanoparticles prepared in this application do not show denaturation or precipitation after heating at 85°C for 40 minutes. Compared with conventional IgG, the IgG nanoparticles prepared in this application have better stability.

[0065] In summary, the thermal and structural stability of immunoglobulin G is significantly enhanced after nano-sizing, making it less prone to denaturation and polymerization at high temperatures. This method also ensures seamless integration of IgG into liquid beverages and other delivery systems without degradation, paving the way for novel, multifunctional biomedical applications. The enhanced properties of IgG nanoparticles make them promising for biomedical applications such as drug delivery and targeted therapy, potentially providing more effective treatments and improving human health. IgG nanoparticles retain and improve their ability to effectively bind antigens, which is crucial for diagnostics, immunotherapy, and vaccine development.

[0066] (2) Detection of denaturation results The immunoglobulin G nanoparticles prepared in Example 1 and conventional IgG without nano-processing were heated at 60℃, 65℃, 70℃, 75℃, 80℃, and 85℃ for 0-120 min, respectively. The heated products were centrifuged, and the percentage of solids in the total volume after centrifugation was calculated and recorded as the denaturation result. The detection results are shown in [Figure 1]. Figure 7 .

[0067] according to Figure 7 It can be seen that there is a significant difference in the thermal stability of immunoglobulin G nanoparticles and conventional IgG without nano-processing.

[0068] Heating immunoglobulin G nanoparticles at 60℃-85℃ for 0-120 minutes did not affect their activity.

[0069] When conventional IgG without nano-processing is heated at 60℃-85℃ for 0-120 minutes, the IgG denatures and precipitates; furthermore, conventional IgG without nano-processing gradually loses its activity as the temperature and heating time increase.

[0070] Therefore, compared with conventional IgG without nano-processing, the IgG nanoparticles prepared by the method of this application maintain good dispersibility, stability and activity, and have an extended half-life, and can be widely used in food, health products, liquid beverages, medical applications, etc.

[0071] With the advancement of nanotechnology, the stability and function of IgG nanoparticles have been significantly enhanced, providing new opportunities for applications in various industries, including the medical field.

[0072] 5. FTIR spectroscopy determination The absorption peaks of the immunoglobulin G nanoparticles prepared in Example 1 and conventional IgG without nano-processing were detected using an infrared spectrometer.

[0073] Infrared spectroscopy revealed the absorption peak characteristics of IgG nanoparticles and conventional IgG samples without nanoparticle processing, such as... Figure 8 As shown, based on the absorption peaks of conventional IgG samples without nano-processing, the NH stretching vibration (amide A) caused a broad and strong absorption band centered at 3300 cm⁻¹. Other strong absorption bands appeared near 1650 cm⁻¹ and 1534 cm⁻¹, caused by the C=O stretching vibration (amide I) and the NH bending vibration (amide II), respectively. The absorption bands centered at 3300 cm⁻¹, 1650 cm⁻¹, 1534 cm⁻¹, and 1075 cm⁻¹ correspond to the protein's NH stretching (amide A), C=O stretching (amide I), NH bending (amide II), and CO stretching vibrations, respectively.

[0074] The spectra of IgG nanoparticles differed significantly from those of conventional IgG without nanoprocessing. Pure IgG nanoparticles without added chemicals exhibited narrower peaks, indicating lower structural variability. In contrast, conventional IgG samples without nanoprocessing showed broader peaks at 3300 cm⁻¹, 1650 cm⁻¹, and 1075 cm⁻¹, reflecting the greater structural stability of the C=O and NH functional groups. Therefore, conventional IgG without nanoprocessing readily binds to other components.

[0075] 1235cm -1 and 1075cm -1 The absorption band at this point corresponds to the antisymmetric stretching vibration of the ester group or the CO stretching of the alcohol, indicating that conventional IgG without nano-processing is more likely to undergo cross-linking reactions with other compounds, forming a polymer structure of immunoglobulin G and other components, i.e., polymerized IgG; while the filament peak (1235 cm⁻¹) -1 and 1075cm -1 The absence of these components in the spectrum of IgG nanoparticles indicates that IgG nanoparticles possess more monomeric, trimeric, or tetrameric structures compared to conventional IgG without nanoprocessing, rather than polymeric structures formed with other components (see [link to original text]). Figure 6 ).

[0076] 6. X-ray diffraction (XRD) measurement X-ray diffraction was used to detect the immunoglobulin G nanoparticles prepared in Example 1 and conventional IgG without nano-processing.

[0077] According to XRD results, the average crystallinity of conventional IgG was 42.52; the average crystallinity of IgG nanoparticles (NPs) was 2.22. The IgG nanoparticles prepared in this invention did not undergo significant polymerization, and their crystallinity was significantly improved compared with conventional IgG without nano-sizing treatment.

[0078] Furthermore, due to the reduced particle size and improved dispersibility, IgG nanoparticles exhibit high stability. XRD pattern (see...) Figure 9 The characteristic diffraction peaks of IgG nanoparticles were observed, demonstrating their good crystalline structure and highly dispersed state.

[0079] 7. Use thermogravimetric analysis (TGA) to determine Thermogravimetric analysis was used to detect the immunoglobulin G nanoparticles prepared in Example 1 and conventional IgG without nano-processing.

[0080] TGA results showed that the weight loss curves of IgG nanoparticles and conventional IgG without nano-processing differed significantly during the heating process. For example... Figure 10 As shown, the weight loss of IgG nanoparticles at 600℃ was 23.7%, while the weight loss of conventional IgG without nano-sizing was 21.8%. This indicates that IgG nanoparticles have high thermal stability, further verifying that they can maintain structural integrity and stability during the preparation process.

[0081] This application discloses an innovative technology for extracting IgG nanoparticles from bovine colostrum. These nanoparticles exhibit strong thermal stability, extended half-life, and enhanced bioactivity. Immunoglobulin G nanoparticles prepared using this method maintain their activity at high temperatures, making them suitable for heat-processed products. The extended half-life reduces the need for frequent ingestion, improving commercial viability. Furthermore, the method employed in this application focuses on preserving and enhancing the antigenic, antibacterial, and immunomodulatory properties of IgG during processing and storage. This method also ensures seamless integration of IgG into liquid beverages and other delivery systems without degradation, paving the way for novel, multifunctional biomedical applications. The enhanced properties of IgG nanoparticles make them promising for biomedical applications such as drug delivery and targeted therapy, potentially providing more effective treatments and improving human health. The IgG nanoparticles retain and enhance their ability to effectively bind antigens, which is crucial for diagnostics, immunotherapy, and vaccine development.

[0082] This application discloses an immunoglobulin G nanoparticle and its industrial preparation method and application. This method fills the gap in the industrial preparation of high-purity immunoglobulin in China. Currently, the purity of immunoglobulin G prepared industrially in China is only 20-30%. The high-purity immunoglobulin G nanoparticles obtained by the method of this invention can reach a purity of up to 80%.

[0083] The findings of this application demonstrate that IgG nanoparticles possess high stability and efficiency, making them promising candidates for biomedical and pharmaceutical applications requiring stable and bioactive formulations.

[0084] IgG nanoparticles maintain structural integrity and amino acid stability under various conditions, making them suitable for biomedical and pharmaceutical applications. Furthermore, the environmentally friendly and cost-effective production method employed in this application highlights the large-scale application potential of IgG nanoparticles in functional foods, diagnostics, and therapeutics. Research could explore encapsulating other bioactive molecules with IgG nanoparticles to create multifunctional nanocarriers. In addition, a thorough examination of the interactions of IgG nanoparticles with various cell types and their immunomodulatory effects is needed to assess their safety and efficacy in clinical settings.

[0085] This study provides valuable insights into the potential industrial applications and versatility of IgG nanoparticles, highlighting their enhanced stability and bioactivity compared to conventional chemical nanoparticles.

Claims

1. An industrial preparation method for immunoglobulin G nanoparticles, characterized in that, The immunoglobulin G nanoparticles were extracted from bovine colostrum, and the preparation method is as follows: (1) The bovine colostrum was centrifuged to remove fat, and the result was defatted bovine colostrum; (2) Heat the skimmed bovine colostrum to 40-50℃, adjust the pH of the skimmed bovine colostrum to 4.4-4.6, filter, remove casein, and obtain whey; (3) Adjust the pH of the whey to 3-3.8, filter, and obtain purified whey; (4) The purified whey was concentrated by ultrafiltration using an ultrafiltration device with a membrane molecular weight cutoff of 10 kDa to obtain nanoparticles; (5) The nanoparticles were dried to obtain immunoglobulin G nanoparticles with a particle size of 10nm-200nm.

2. The industrial preparation method of immunoglobulin G nanoparticles according to claim 1, characterized in that: In step (1), the temperature of the bovine colostrum is controlled at 4-18℃ before centrifugation and defatting. When performing centrifugation for defatting in step (1), the centrifugation speed is 3000-6000 rpm, and the temperature of bovine colostrum is controlled to be ≤25℃ during the centrifugation process.

3. The industrial preparation method of immunoglobulin G nanoparticles according to claim 1, characterized in that: In step (2), the solution used to adjust the pH of the skimmed bovine colostrum is an acidic solution; in step (3), the solution used to adjust the pH of the whey is an acidic solution.

4. The industrial preparation method of immunoglobulin G nanoparticles according to claim 3, characterized in that: The acid solution is hydrochloric acid solution, citric acid solution, vitamin C solution, or acetic acid solution.

5. The industrial preparation method of immunoglobulin G nanoparticles according to claim 1, characterized in that: In step (2), the filtration process is as follows: the pH-adjusted skimmed bovine colostrum is filtered through a 60-200 mesh sieve.

6. The industrial preparation method of immunoglobulin G nanoparticles according to claim 1, characterized in that, In step (3), the filtration process is as follows: the whey after pH adjustment is filtered through a sieve with a mesh size of ≥200.

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