Macroporous bovine serum albumin nanoparticles as well as preparation method and application thereof

Abstract

The invention relates to a macroporous bovine serum albumin nanoparticle and a preparation method and application thereof, the macroporous bovine serum albumin nanoparticle is a nanoparticle with macropores distributed on the surface, the size range of the nanoparticle is 300-645nm, and the macropores are through holes with the aperture range of 50-100nm; the nanoparticles are symmetrical N-surface or spherical nanoparticles, wherein N is greater than or equal to 12. The specific surface area reaches up to 20-60 m / g, the drug loading capacity (5-25% w / w) and the encapsulation efficiency (50-90%) are remarkably improved, and the adsorption efficiency is greatly improved compared with that of traditional nanoparticles without holes in the surfaces. Compared with BSA nano-particles without holes in the surface, the absorption efficiency of RhB is improved due to the existence of macroholes, and the BSA nano-particles have wide application in the aspect of drug delivery.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and in particular to macroporous bovine serum albumin nanoparticles, a preparation method thereof, and applications thereof. Background Art

[0002] Bovine serum albumin nanoparticles (BSA nanoparticles or BNP) are biodegradable, biocompatible, nontoxic, and non-immunogenic, offering advantages over synthetic polymers in medical applications such as drug delivery. However, existing BSA nanoparticles are typically prepared using co-precipitation methods. This process is uncontrollable, resulting in a wide and uneven size distribution of the resulting nanoparticles. Furthermore, impurities may be introduced during precipitation and subsequent processing, affecting the purity of the nanoparticles. Furthermore, the loading capacity of the resulting BSA nanoparticles needs to be further improved. Summary of the Invention

[0003] The object of the present invention is to provide a macroporous bovine serum albumin nanoparticle capable of obtaining uniform particles and good attachment ability, as well as a preparation method and use thereof.

[0004] A macroporous bovine serum albumin nanoparticle having macropores distributed on its surface. The nanoparticle has a size range of 300-645 nm, and the macropores are through-holes with a pore size range of 50-100 nm. The nanoparticle is a symmetrical N-hedron or spherical nanoparticle, wherein N is ≥ 12. The specific surface area of the nanoparticle is 20-60 m² / g.

[0005] Furthermore, the adsorption efficiency of the nanoparticles of the present invention is 5-20 mg / g. The adsorption efficiency can be expressed by the amount of target molecules adsorbed per unit mass of the particles, that is, the number of milligrams of molecules adsorbed per gram of particles.

[0006] A method for preparing macroporous bovine serum albumin nanoparticles based on any of the above-mentioned methods, wherein the method is a method for preparing macroporous bovine serum albumin nanoparticles based on a vortex fluid device, comprising the following steps:

[0007] 1) First, an ethanol solution is added to bovine serum albumin for desolvation to obtain a raw material solution, and then the raw material solution is combined with an appropriate amount of ethanol and an appropriate amount of glutaraldehyde to form a solution to be treated;

[0008] 2) Transfer the solution to be treated to an inclined vortex fluid device and rotate it at 6000-7500 rpm for 1-1.5 minutes to obtain a treated solution;

[0009] 3) The treated liquid is centrifuged and washed to obtain the macroporous bovine serum albumin nanoparticles.

[0010] The rotation time of the solution to be treated in the vortex fluid device should not be too long or too short. If the treatment time is too short, the diameter of the nanoparticles is generally larger. Increasing the treatment time will further disperse the nanoparticles, forming smaller particles. However, too long a treatment time may cause the nanoparticles to re-aggregate, resulting in larger nanoparticles and the formation of irregular pocket-shaped structures.

[0011] A suitable rotational speed not only produces nanoparticles of appropriate size, but also allows the nanoparticles to be stably shaped into N-hedrons or spheres. If the rotational speed is too slow, the shear force is relatively reduced, and localized stress concentration on the nanoparticle surface may cause the nanoparticles to form irregular "pockets" or irregularly shaped structures. Conversely, if the rotational speed is too high, the nanoparticles are too small.

[0012] Furthermore, the vortex fluid device of the preparation method of the present invention includes a working unit, the working unit and the horizontal direction have an inclination angle of 40~50 °, the outer diameter of the vortex fluid device is 10~30mm, the inner diameter is 17.1~17.5mm, and the length is 19.4cm~39cm. The appropriate inclination angle can provide a moderate gravity effect, making the movement of the fluid in the rotating tube more stable and controllable, both avoiding excessive liquid aggregation and effectively controlling the distribution of shear force. If the inclination angle is less than 40 °, the shear force and vortex effect will be significantly weakened, and the flow pattern of the fluid may not be able to provide enough mixing and shearing effect, which will reduce the generation efficiency of the macroporous structure. But on the contrary, if it exceeds 50 °, it may cause the gravity to be too large, and the liquid cannot form a stable film in the tube.

[0013] Furthermore, the vortex fluid device of the preparation method of the present invention also includes a tilting frame, a rotating system arranged on the tilting frame and a working unit connected to the rotating system, as well as a liquid inlet and discharge system, a centrifugal and recovery system; the working unit is a glass tube.

[0014] Furthermore, the concentration of the ethanol solution in the preparation method of the present invention ranges from 0.5 to 2 mg / L, the concentration of bovine serum albumin in the raw material solution ranges from 0.5 to 2 mg / L, and the volume ratio of bovine serum albumin:ethanol:glutaraldehyde in the treated solution is (15-25):(55-65):(0.5-1). Ethanol influences the size of the nanoparticles by affecting particle aggregation and dispersion during the desolvation process. If the ethanol ratio is too high, the solubility of BSA decreases, resulting in very small nanoparticles. Conversely, if the ethanol ratio is too low, the driving force provided is too low, the aggregation rate of BSA is slow, and the particles are more likely to form holes and irregular shapes in localized areas, resulting in pores or even "pocket" structures on the particle surface. The glutaraldehyde concentration affects the degree of cross-linking on the particle surface and the compactness of the particle internal structure, which in turn affects the diameter. If the glutaraldehyde concentration is too low, the nanoparticles are not sufficiently cross-linked, and localized areas of loose structure may form, resulting in "pocket" shapes or irregular morphologies on the nanoparticle surface.

[0015] Furthermore, the volume ratio of the solution to be treated to the inner cavity of the glass tube in the preparation method of the present invention is (0.3-0.6):1. Too much solution to be treated in the glass tube will result in oversized and irregularly shaped nanoparticles; too little will reduce efficiency.

[0016] Furthermore, the centrifugation and washing process in step 3) of the preparation method of the present invention specifically includes: removing excess reactants by centrifugation at 10,000-12,000 g for 10-20 min and washing with the same volume of Milli Q water at least three times.

[0017] A use of the macroporous bovine serum albumin nanoparticles according to any one of the above items, wherein the macroporous bovine serum albumin nanoparticles are used for drug delivery.

[0018] Furthermore, when used for drug delivery, the macroporous bovine serum albumin nanoparticles described herein have a drug loading of 5-25% and an encapsulation efficiency of 50-90%. Drug loading is expressed as weight percent (w / w%), i.e., the percentage of drug mass loaded into the nanoparticles relative to the total mass of the particles. Encapsulation efficiency, typically expressed as a percentage, indicates the proportion of drug added to the system that is successfully encapsulated by the nanoparticles.

[0019] Compared with the prior art, the present invention has the following beneficial technical effects:

[0020] The macroporous bovine serum albumin nanoparticles and their preparation method described in the present invention utilize a vortex fluid device to precisely control fluid shear force and the desolvation process, thereby preparing, for the first time, BSA nanoparticles with 50-100 nm through-hole macropores and a symmetrical N-hedral / spherical structure (N ≥ 12). Their specific surface area reaches 20-60 m² / g, significantly improving drug loading capacity (5-25% w / w) and encapsulation efficiency (50-90%), significantly enhancing adsorption efficiency compared to conventional nanoparticles with non-porous surfaces.

[0021] Moreover, the working unit of the vortex fluid device used has an inclination angle of 40-50° with the horizontal direction. The outer diameter of the vortex fluid device is 10-30 mm, the inner diameter is 17.1-17.5 mm, and the length is 19.4 cm-39 cm. The appropriate inclination angle can provide a moderate gravity effect, making the movement of the fluid in the rotating tube more stable and controllable, avoiding excessive liquid aggregation and effectively controlling the distribution of shear force. By limiting the vortex rotation speed (6000-7500 rpm), processing time (1-1.5 min), device inclination angle (40-50°) and solution ratio (BSA: ethanol: glutaraldehyde = 15-25: 55-65: 0.5-1), precise control of nanoparticle size (300-645 nm) and pore structure is achieved, overcoming the problems of wide particle distribution, easy aggregation and impurity residue in traditional methods.

[0022] Furthermore, the present invention utilizes a short centrifugation (10-20 minutes, 10,000-12,000g) combined with three pure water washes to effectively remove residual glutaraldehyde, ensuring the nanoparticles' non-toxicity and non-immunogenicity, thus meeting safety requirements for drug delivery. Furthermore, the macroporous through-hole structure of the present invention facilitates drug loading and its symmetrical morphology, enhancing in vivo circulation stability. This provides a highly efficient and controllable nanocarrier platform for medical applications such as tumor targeting and gene therapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Schematic diagram of the structure of the vortex fluid device of the present invention;

[0024] Figure 2 This is a microscopic morphology of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention;

[0025] Figure 3 This is a microscopic morphology of the sample obtained in Comparative Example 2 of the present invention;

[0026] Figure 4 Fluorescence microscopy analysis of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention;

[0027] Figure 5UV-Vis and fluorescence spectral analysis of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention; blue: fluorescence emission at an excitation wavelength of 490 nm; red: absorption;

[0028] Figure 6 To characterize the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention using confocal imaging;

[0029] Figure 7 is Figure 6 The emission wavelength of a single macroporous BNP highlighted by a colored circle in the figure, where the sample was excited with a laser at a wavelength of 532 nm, and the blue one is the macroporous BSA nanoparticles;

[0030] Figure 8 CD analysis of bovine serum albumin and macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention; the blue ones are macroporous bovine serum albumin nanoparticles;

[0031] Figure 9 This is the nitrogen adsorption analysis of the porous BNPs of bovine serum albumin and the prepared macroporous bovine serum albumin nanoparticles in Example 1 of the present invention; the blue ones are macroporous bovine serum albumin nanoparticles;

[0032] Figure 10 FTIR analysis of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention;

[0033] Figure 11 This is a Raman analysis of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention;

[0034] Figure 12 This is the XRD analysis of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention;

[0035] Figure 13 STA analysis of the macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention;

[0036] Figure 14 The fluorescence spectrum of RhB loaded with macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention is studied;

[0037] Figure 15 Fluorescence microscopy of the RhB-loaded macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention under an Invitrogen EVOS light cube for bright field, GFP (470 / 22 Ex; 510 / 42 Em), and RFP (531 / 40 Ex; 593 / 40 Em), respectively;

[0038] Figure 16To characterize BNP / CPC using SEM, smooth surface, less porous surface, and vesicle-like morphology are shown from left to right. DETAILED DESCRIPTION

[0039] In order to have a clearer understanding of the technical features, purposes and effects of the present invention, the specific embodiments of the present invention are now described in detail with reference to the accompanying drawings. The described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. If specific conditions are not specified in the specific implementation method, they shall be carried out according to conventional conditions or conditions provided by the manufacturer.

[0040] When the following description refers to the accompanying drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present disclosure. The terms used in this disclosure are for the purpose of describing specific embodiments only and are not intended to limit the present disclosure. Specific implementation method one:

[0042] A macroporous bovine serum albumin nanoparticle having macropores distributed on its surface, wherein the nanoparticle has a size range of 300-645 nm, and the macropores are through-holes with a pore size range of 50-100 nm. The nanoparticle is a symmetrical N-hedron or spherical nanoparticle, wherein N is ≥ 12, and the specific surface area of the nanoparticle is 20-60 m² / g.

[0043] In some other embodiments, the adsorption efficiency of the nanoparticles of the present invention is 5-20 mg / g. Specific implementation method two:

[0045] A method for preparing macroporous bovine serum albumin nanoparticles according to any of the above embodiments, wherein the method is a method based on a vortex fluid device.

[0046] The steps include:

[0047] 1) First, an ethanol solution is added to bovine serum albumin for desolvation to obtain a raw material solution, and then the raw material solution is combined with an appropriate amount of ethanol and an appropriate amount of glutaraldehyde to form a solution to be treated;

[0048] 2) Transfer the solution to be treated to an inclined vortex fluid device and rotate it at 6000-7500 rpm for 1-1.5 minutes to obtain a treated solution;

[0049] 3) The treated liquid is centrifuged and washed to obtain the macroporous bovine serum albumin nanoparticles.

[0050] In other embodiments, the concentration range of the ethanol solution in the preparation method is 0.5~2 mg / L, the concentration range of bovine serum albumin in the raw material solution is 0.5~2 mg / L, and the volume ratio of bovine serum albumin: ethanol: glutaraldehyde in the solution to be treated is (15~25): (55~65): (0.5~1).

[0051] In other embodiments, the volume ratio of the solution to be treated in the preparation method to the inner cavity of the glass tube is (0.3-0.6):1.

[0052] In other embodiments, the centrifugation and washing process in step 3) of the preparation method specifically includes: removing excess reactants by centrifugation at 10,000-12,000 g for 10-20 min and washing with the same volume of Milli Q water at least three times. Specific implementation method three:

[0054] A vortex fluid device comprises a working unit, a tilting frame, a rotating system mounted on the tilting frame, a working unit connected to the rotating system, a liquid inlet and outlet system, and a centrifugal and recovery system. In this embodiment, the working unit is a glass tube. The working unit is tilted at an angle of 40 to 50 degrees relative to the horizontal. The vortex fluid device has an outer diameter of 10 to 30 mm, an inner diameter of 17.1 to 17.5 mm, and a length of 19.4 to 39 cm.

[0055] The rotation system can adjust the speed. The rotation speed and duration are key parameters that influence the formation and structural properties of nanoparticles. The raw material solution is introduced into the glass tube via the inlet system, typically using a desolvation method. During the vortexing process, the components in the solution are spun to form a thin film and mixed at high speed. After the treatment, the resulting sample is recovered by centrifugation.

[0056] The vortex fluid device produces a vortex fluid film by the glass tube of high-speed rotation. In this film, reactants are strongly sheared and mixed to form an efficient fluid dynamic environment. This shearing force can not only accelerate the reaction process, but also generate macroporous nanoparticles with high uniformity. By accurately controlling the ratio of rotational speed, tilt angle, reactants and the processing time, the size and shape of nanoparticles can be regulated. In addition, the film fluid dynamics of the vortex fluid device can also accelerate the synthesis of protein folding, nanomaterial peeling and drug delivery carriers.

[0057] In addition, the generated vortex fluid film can provide good mixing and diffusion conditions, thereby promoting the formation of nanoparticles and effectively controlling the morphology of the nanoparticles, especially the generation of pores. The highly uniform shear force field can form uniform large pores on the surface of the generated nanoparticles. Specific implementation method four:

[0059] A use of the macroporous bovine serum albumin nanoparticles according to any of the above specific embodiments, wherein the macroporous bovine serum albumin nanoparticles are used for drug delivery.

[0060] When the macroporous bovine serum albumin nanoparticles are used for drug delivery, the drug loading amount is 5-25% and the encapsulation efficiency is 50-90%.

[0061] The present invention is further described in detail below with reference to specific embodiments.

[0062] Example 1:

[0063] A method for preparing macroporous bovine serum albumin nanoparticles, the preparation method is a preparation method based on a vortex fluid device. In this embodiment 1, as Figure 1 As shown, the vortex fluid device includes a working unit, a tilting frame, a rotating system provided on the tilting frame, a glass tube connected to the rotating system, as well as a liquid inlet and discharge system, a centrifugal and recovery system; wherein the glass tube is tilted at an angle of 45° to the horizontal direction, and the vortex fluid device has an outer diameter of 20 mm, an inner diameter of 17.1 mm, and a length of 19.4 cm.

[0064] The preparation method comprises the following steps:

[0065] 1) First, an ethanol solution is added to bovine serum albumin for desolvation to obtain a raw liquid, wherein the concentration of the ethanol solution is 1 mg / L, and the concentration of bovine serum albumin in the raw liquid is within a range of 1 mg / L. The raw liquid is then combined with an appropriate amount of ethanol and an appropriate amount of glutaraldehyde to form a solution to be treated; the volume ratio of bovine serum albumin:ethanol:glutaraldehyde in the solution to be treated is (15-25):(55-65):(0.5-1). The volume ratio of the solution to be treated to the inner lumen of the glass tube is (0.3-0.6):1. In this Example 1, the volume ratio of bovine serum albumin to ethanol is 1:2.

[0066] 2) Transfer the solution to be treated to an inclined vortex fluid device and rotate it at 6000 rpm for 1 min to obtain a treated solution;

[0067] 3) The treated solution was centrifuged and washed to obtain the macroporous bovine serum albumin nanoparticles, which were centrifuged at 11800 g for 15 min and washed three times with the same volume of Milli Q water to remove excess reactants.

[0068] Milli-Q water is ultrapure water widely used in laboratories and scientific research. Milli-Q water quality standards: Milli-Q water meets Class I water quality standards established by ASTM, CAP, NCCL, EP, and USP, with a resistivity of 18.2 MΩ·cm, a total organic carbon (TOC) content of less than 5 ppm, a bacterial content of less than 0.1 cfu / mL, an endotoxin content of less than 0.001 EU / mL, and extremely low levels of ribonucleases (RNases) and deoxyribonucleases (DNases).

[0069] In all the drawings of the present invention, BNP and BNP-HP are macroporous bovine serum albumin nanoparticles prepared in Example 1; BSA is the raw material bovine serum albumin in Example 1.

[0070] like Figure 2 、 Figure 4 and Figure 5 Shown are the microscopic morphology, fluorescence microscopy analysis, and UV-Vis and fluorescence spectroscopy analysis of macroporous bovine serum albumin nanoparticles.

[0071] It can be seen that the macroporous bovine serum albumin nanoparticles are nanoparticles with macropores distributed on the surface, and the size range of the nanoparticles is 300-645 nm.

[0072] like Figures 6 to 9 As shown, the difference between the nanoparticles prepared in Example 1 and Comparative Example 1 was explored by confocal imaging:

[0073] As can be seen from the figure, randomly distributed macropores are displayed in the spherical particles, and the pore size of the through-type macropores ranges from 50 to 100 nm. Some of the particles are approximately dodecahedral symmetric, with a macropore on each pentagonal face. Confocal mapping was performed on BNP, which allows direct measurement of single particle emission at about 555 nm after excitation at 532 nm. This indicates the denaturation of the raw bovine serum albumin received and the formation of new chromophores. In combination with CD, it was clearly determined that in the preparation method described in the present application, BSA underwent significant structural changes during the preparation. BSA mainly contains α-helix as a secondary structural element, which produces two negative bands at 208 and 220 nm in the CD spectrum, as shown in the figure. Figure 8 The conformation of BNP is thought to be different from the native conformation of BSA in aqueous solution. BNP forms an emulsion with two major peaks near 207 and 235 nm in the CD spectrum, which may indicate denaturation of the α-helical structure into β-sheets.

[0074] Hydrogen bonding interactions can bring together antiparallel peptide chains in adjacent BSA molecules, generating a β-sheet structure that perturbs the CD spectrum. These results highlight the modification of the BSA α-helical secondary structure during treatment in the vortex fluidics device, consistent with protein reorganization to form a new chromophore.

[0075] It should be noted that Figure 7 The red and blue circles are shown in the figure, which clearly mark the "strongest emission wavelength" of the two curves. These two circles make it easy to compare the peak position differences between the sample (red) and macroporous bovine serum albumin nanoparticles (blue), and then deduce the differences in the composition, structure or optical properties of the nanoparticles.

[0076] The differences between the nanoparticles prepared in Example 1 and Comparative Example 1 were investigated by X-ray diffraction patterns:

[0077] Depend on Figures 10-13 As shown, BNP has the peaks at 3306, 1657, and 1539 cm -1 The FTIR absorption peaks at correspond to OH stretching vibration, amide I band (mainly CO stretching vibration) and amide II band (mainly NH and CN). The absence of Raman features may be due to the fluorescence of the sample under 532 nm laser irradiation, such as Figure 11 X-ray diffraction (XRD) highlighted the amorphous nature of the as-received BSA and macroporous BNPs, with slight differences at 2θ = 12°, as shown in Figure 12 The stable amorphous form is advantageous in having a higher dissolution rate, which enhances drug release and subsequent absorption and bioavailability. STA analysis of BNP and native BSA showed no significant difference. Figure 13 Both samples begin to degrade at approximately 250°C, followed by a sudden weight loss at approximately 300°C, likely due to the loss of small molecules such as carbon dioxide and ammonia. A slight difference in weight loss between the two is observed between 350 and 550°C, with the difference slowly increasing above 500°C. This is likely due to the reduced crystalline nature of BNP relative to as-prepared BSA, as shown in XRD.

[0078] The differences between the nanoparticles prepared in Example 1 and Comparative Example 1 were investigated by potential difference and fluorescence microscopy:

[0079] BNP remains stable under physiological conditions. The potential difference shows that the positively charged RhB interacts with the negatively charged BSA, promoting the binding of RhB to BNP. Due to the adsorption of RhB by BNP, the concentration of RhB gradually decreases over time, and the adsorption amount of RhB by BNPs with different morphologies is different. BNP-HP exhibits the highest loading capacity. RhB is 1.44 nm × 1.09 nm × 0.64 nm and can penetrate micropores with a pore size of > 1.5 nm. In Comparative Example 1, in the absence of pores, adsorption will only occur on the outer surface of the particles, so a lower amount of RhB is loaded. The fluorescence spectrum provides characteristic peaks of RhB (Ex 550 nm) and BNP (Ex 490 nm). Compared with BNP with other surface morphologies, the fluorescence intensity of macroporous BNP-RhB is significantly enhanced, as shown in Figure 2. Figure 14 As shown. Figure 15 , which are fluorescence microscopy images of the RhB-loaded macroporous bovine serum albumin nanoparticles prepared in Example 1 of the present invention under an Invitrogen EVOS light cube, for bright field, GFP (470 / 22 Ex; 510 / 42 Em) and RFP (531 / 40 Ex; 593 / 40 Em), respectively.

[0080] Comparative Example 1:

[0081] In this comparative example 1, nanoparticles were prepared using bovine serum albumin as a raw material. The specific process was different from that of Example 1 only in that a desktop vortex treatment was used instead of the vortex fluid device in Example 1.

[0082] Comparative Example 2:

[0083] The only difference between Comparative Example 2 and Example 1 is that the outer diameter of the vortex fluid device is 20 mm, the inner diameter is 17.1 mm, and the length is 19.1 cm. Finally, the following is obtained: Figure 3 Samples shown.

[0084] Comparative Example 3:

[0085] Comparative Example 3 differs from Example 1 only in that the treated solution also includes C-phycocyanin. Consequently, the resulting macroporous BSA nanoparticles exhibit surface features that are not perfectly regular spheres but rather have concave or "pocket-like" characteristics. This is because the addition of C-phycocyanin alters the aggregation and cross-linking patterns of BSA during its binding process.

[0086] like Figure 16As shown, the size of BNP particles can be easily reduced by 400 by incorporating another fluorescent protein called C-PC (Ex 620 nm, Em 650 nm) into the treatment of BSA. The α (162 residues) and β (172 residues) subunits of C-PC form a heterodimeric αβ monomer, which contains three chromophores, one in the α subunit and two in the β subunit, and the three monomers can oligomerize to form a discoid trimer (αβ)3. Figure 16 Shows the use of Figure 3 The shape and size of the newly obtained BNP (as BNP / CPC) were determined under the same experimental conditions as for BSA alone, with the only difference being the addition of C-PC. A noticeable difference in the composite protein particles is the significant reduction in size from approximately 100 microns, from 358 nm to 150 nm. Furthermore, instead of forming pores on the surface, pocket-like structures, such as Figure 16 shown.

[0087] The present invention is described by the above-mentioned specific embodiments. It should be understood by those skilled in the art that various changes and equivalent substitutions may be made to the present invention without departing from the scope of the present invention. Parts not described in detail in the present specification are well-known technologies to those skilled in the art. In addition, various modifications may be made to the present invention for specific situations or specific circumstances without departing from the scope of this new use. Therefore, the present invention is not limited to the specific embodiments disclosed, but should include all embodiments falling within the scope of the present invention.

Claims

1. A macroporous bovine serum albumin nanoparticle, characterized in that: The macroporous bovine serum albumin nanoparticles are nanoparticles with macropores distributed on the surface. The size of the nanoparticles ranges from 300 to 645 nm, and the macropores are through-holes with a pore size range of 50 to 100 nm. The nanoparticles are symmetrical N-hedron or spherical nanoparticles, where N is ≥ 12. The specific surface area of the nanoparticles is 20 to 60 m² / g.

2. The macroporous bovine serum albumin nanoparticles according to claim 1, characterized in that The adsorption efficiency of the nanoparticles is 5-20 mg / g.

3. A method for preparing macroporous bovine serum albumin nanoparticles according to any one of claims 1 or 2, characterized in that: The preparation method is a preparation method based on a vortex fluid device, The steps include: 1) First, an ethanol solution is added to bovine serum albumin for desolvation to obtain a raw material solution, and then the raw material solution is combined with an appropriate amount of ethanol and an appropriate amount of glutaraldehyde to form a solution to be treated; 2) Transfer the solution to be treated to an inclined vortex fluid device and rotate it at 6000-7500 rpm for 1-1.5 minutes to obtain a treated solution; 3) The treated liquid is centrifuged and washed to obtain the macroporous bovine serum albumin nanoparticles.

4. The preparation method according to claim 3, characterized in that The vortex fluid device includes a working unit, the working unit is inclined at an angle of 40-50° to the horizontal direction, the outer diameter of the vortex fluid device is 10-30 mm, the inner diameter is 17.1-17.5 mm, and the length is 19.4 cm-39 cm.

5. The preparation method according to claim 4, characterized in that The vortex fluid device also includes a tilting frame, a rotating system arranged on the tilting frame, a working unit connected to the rotating system, as well as a liquid inlet and outlet system, a centrifugal and recovery system; the working unit is a glass tube.

6. The preparation method according to claim 5, characterized in that The concentration range of the ethanol solution is 0.5-2 mg / L, the concentration range of bovine serum albumin in the raw material solution is 0.5-2 mg / L, and the volume ratio of bovine serum albumin: ethanol: glutaraldehyde in the solution to be treated is (15-25): (55-65): (0.5-1).

7. The preparation method according to claim 6, characterized in that The volume ratio of the solution to be treated to the inner cavity of the glass tube is (0.3-0.6):

1.

8. The preparation method according to claim 7, characterized in that The centrifugation and washing process in step 3) specifically includes: removing excess reactants by centrifugation at 10,000-12,000 g for 10-20 min and washing with the same volume of Milli Q water at least three times.

9. A use of the macroporous bovine serum albumin nanoparticles according to any one of claims 1 to 8, characterized in that: The macroporous bovine serum albumin nanoparticles are used for drug delivery.

10. The use according to claim 9, characterized in that When the macroporous bovine serum albumin nanoparticles are used for drug delivery, the drug loading amount is 5-25% and the encapsulation efficiency is 50-90%.

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