CN109730975B - A method for maintaining the biological activity of drug-loaded erythrocytes while increasing their drug-loading capacity - Google Patents
A method for maintaining the biological activity of drug-loaded erythrocytes while increasing their drug-loading capacity Download PDFInfo
- Publication number
- CN109730975B CN109730975B CN201811559344.0A CN201811559344A CN109730975B CN 109730975 B CN109730975 B CN 109730975B CN 201811559344 A CN201811559344 A CN 201811559344A CN 109730975 B CN109730975 B CN 109730975B
- Authority
- CN
- China
- Prior art keywords
- drug
- loaded
- red blood
- glucan
- blood cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明属于药物制剂领域，具体涉及一种维持载药红细胞生物活性同时提高载药量的方法。所述方法至少包括以下步骤：向低渗葡聚糖药液中加入红细胞，第一次孵育；加入高渗液，第二次孵育，使低渗葡聚糖药液中的目的药物包封于红细胞中，离心洗涤除去葡聚糖，得到载药红细胞。采用本发明提供的方法简便，高效，效果好，最后制得的载药红细胞的各类生物活性指标与天然红细胞没有显著性差异，很好地达到了维持载药红细胞体外生物活性的目的；在维持生物活性的同时，本发明提供的方法还能提高药物的包载量。The invention belongs to the field of pharmaceutical preparations, and in particular relates to a method for maintaining the biological activity of drug-carrying red blood cells while increasing the drug-carrying amount. The method includes at least the following steps: adding red blood cells to the hypotonic glucan liquid, incubating for the first time; adding hypertonic liquid, and incubating for the second time, so that the target drug in the hypotonic glucan liquid is encapsulated in the hypotonic glucan liquid. In the erythrocytes, the glucan was removed by centrifugation and washing to obtain drug-loaded erythrocytes. The method provided by the invention is simple, efficient and effective, and the various biological activity indexes of the finally prepared drug-loaded red blood cells are not significantly different from those of natural red blood cells, and the purpose of maintaining the in vitro biological activity of the drug-loaded red blood cells is well achieved; While maintaining the biological activity, the method provided by the present invention can also improve the encapsulation capacity of the drug.
Description
Technical Field
The invention belongs to the field of pharmaceutical preparations, and particularly relates to a method for maintaining the biological activity of drug-loaded red blood cells and improving the drug-loaded amount of the drug-loaded red blood cells.
Background
The Red Blood Cell (RBC) is the most abundant cell type of human body, about one fourth of the total cell number, and the mature red blood cell does not contain organelles, nucleic acid and other substances, and the content components are simple. The circulation time of the red blood cells in vivo is long, wherein the human RBC can survive for 100-120 days after maturation and the rat red blood cells can survive for 50-60 days. Based on the characteristics of erythrocytes, i.e., abundant sources, good biocompatibility, good biodegradability, no induction of immune response, long in vivo circulation time, etc., erythrocytes have been widely used for entrapping various substances, including enzymes, nucleic acids and various small molecule drugs, since the last 60-70 th century. However, when the erythrocytes completing the in vitro drug loading process re-enter the circulation in vivo, the service life of the erythrocytes is often only 1/3 of natural erythrocytes or even shorter, and no solution for solving the problems is available at present.
Disclosure of Invention
The invention aims to provide a method for maintaining the biological activity of drug-loaded red blood cells and simultaneously improving the drug-loading rate of the drug-loaded red blood cells, which can protect the red blood cells from being influenced by various in vitro adverse factors to the greatest extent so as to reduce the biological activity and simultaneously improve the drug-loading rate. In order to achieve the above objects and other related objects, the present invention adopts the following technical solutions:
in a first aspect of the present invention, a method for maintaining the bioactivity of a drug-loaded red blood cell and simultaneously increasing the drug-loading rate of the drug-loaded red blood cell is provided, which at least comprises the following steps:
(1) adding red blood cells into the hypotonic dextran liquid medicine, and incubating for the first time;
(2) and (2) adding hypertonic solution into the mixture incubated in the step (1), incubating for the second time to encapsulate the target drug in the hypotonic dextran liquid medicine into the red blood cells, and centrifuging and washing to remove dextran to obtain the drug-loaded red blood cells.
In the step (1), the hypotonic dextran solution is a hypotonic aqueous solution containing dextran, sodium chloride and a target drug to be loaded on erythrocytes.
The dextran is water soluble. Optionally, the dextran is water soluble.
The molecular weight of the glucan is 10000-70000.
In one embodiment, the hypotonic dextran solution has a mass fraction of dextran of 2% to 20% based on the total amount of the hypotonic dextran solution.
In one embodiment, the hypotonic dextran solution comprises the following components based on the total amount of the hypotonic dextran solution:
the target medicine is 1mg/mL-10mg/mL,
sodium chloride with the mass fraction of 0.4 to 0.6 percent,
2 to 20 percent of glucan by mass,
the solvent is water.
In one embodiment, the drug of interest is selected from the group consisting of betamethasone sodium phosphate, dexamethasone sodium phosphate, doxorubicin hydrochloride, sodium artesunate, morphine hydrochloride, and bovine serum albumin.
In one embodiment, in step (1), the red blood cells are polymerized with hypotonic dextran
The volume ratio of the sugar liquid medicine is 1 (4-20).
In one embodiment, in step (1), the red blood cells are packed red blood cells.
Further, the preparation method of the packed red blood cells at least comprises the following steps: and (3) centrifugally separating animal whole blood, removing plasma and white blood cells, and washing to obtain the packed red blood cells.
Animal whole blood centrifugation conditions were: 4 ℃ and 3800 r/min.
The animal is a mammal, and can be a rodent or a primate. Examples of the compounds include monkeys, rabbits, goats, and rats.
In one embodiment, in step (1), the temperature of the first incubation is: 0 to 10 ℃. Optionally, from 0 to 4 ℃.
In one embodiment, in step (1), the time for the first incubation is 10-60 min.
Further, in the step (2), the hypertonic solution comprises the following components by taking the total amount of the hypertonic solution as a reference: the mass fraction is 2.5-5.5% sodium chloride, 1-10mg/ml sodium pyruvate and 2-20mg/ml glucose, and the solvent is water.
In one embodiment, in step (2), the volume ratio of the hypertonic solution to the hypotonic dextran solution is 1 (4-20).
In one embodiment, in step (2), the temperature of the second incubation is: 30-40 ℃.
In one embodiment, in step (2), the time for the second incubation is 10-60 min.
In a second aspect of the invention, there is provided the use of the aforementioned method for the preparation of a drug-loaded red blood cell preparation.
Compared with the prior art, the invention has the following beneficial effects:
1) the method provided by the invention is simple, convenient, efficient and good in effect, various biological activity indexes of the finally prepared drug-loaded red blood cells have no significant difference with natural red blood cells, and the aim of maintaining the in vitro biological activity of the drug-loaded red blood cells is well fulfilled.
2) The method provided by the invention can improve the entrapment quantity of the medicine while maintaining the biological activity.
Drawings
Other features, objects and advantages of the invention will become more apparent upon reading of the detailed description of non-limiting embodiments with reference to the following drawings:
FIG. 1 is a graph of an experiment for examining the influence of dextran with different molecular weights and different concentrations on the drug loading of erythrocytes;
FIG. 2 is an experimental chart for investigating the influence of dextran with different molecular weights and different concentrations on the osmotic fragility of drug-loaded erythrocytes;
FIG. 3 is an experimental graph for examining the influence of different molecular weights and concentrations of glucan on the Na +/K + -ATPase activity of drug-loaded erythrocytes;
FIG. 4A is a graph of an experiment examining the effect of dextran on phosphatidylserine eversion on drug-loaded erythrocyte membranes; FSC-SSC flow chart of natural erythrocyte Negative Control group (Negative Control);
FIG. 4B is a graph of an experiment examining the effect of dextran on phosphatidylserine eversion on drug-loaded erythrocyte membranes; is the FSC-SSC flow diagram of Natural Red Blood Cells (NRBC);
FIG. 4C is an experimental diagram for examining the effect of dextran on phosphatidylserine eversion on drug-loaded erythrocyte membranes; FSC-SSC flow diagrams for drug-loaded red blood cells (BSP-RBC) prepared without dextran;
FIG. 4D is a graph showing the effect of dextran on phosphatidylserine eversion on the membrane of drug-loaded erythrocyte; is an FSC-SSC diagram of a drug-loaded erythrocyte prepared after adding glucan (T40-10%, wherein T40 represents the molecular weight of the glucan 40000, and 10% represents the mass fraction of the glucan);
FIG. 4E is a graph showing the effect of dextran on phosphatidylserine eversion on drug-loaded erythrocyte membranes; is a diagram of the results of phosphatidylserine eversion experiments.
FIG. 5 is an experimental diagram for investigating the influence of dextran on the morphological structure of drug-loaded erythrocytes; wherein: 5A is NRBC, 5B is BSP-RBC, and 5C is T40-10% -BSP-RBC.
Representative in each figure, p < 0.05, showing significant differences compared to BSP-RBC group; # represents, p < 0.05, showing significant difference compared to the NRBC group. (Spss software paired t-test).
Detailed Description
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the respective manufacturers.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts.
Example 1 addition of dextran and preparation of corresponding drug-loaded erythrocytes
Experimental group (addition of dextran):
(1) a male SD rat with a weight of about 200g is subjected to blood sampling of the posterior orbital venous plexus by a capillary with a diameter of 0.5mm, and the blood sampling amount is 0.2-0.3ml each time. Centrifuging at 4 deg.C and 2000r/min for 4min, discarding supernatant, removing plasma and leukocyte layer, and washing with 1ml pre-cooled physiological saline for 1-2 times until the supernatant is nearly colorless to obtain lower layer packed red blood cells.
(2) Adding 100 μ l of washed erythrocyte into 400 μ l of hypotonic dextran solution, mixing, standing at 4 deg.C for 20min, and shaking gently every 4 min. Wherein the hypotonic dextran liquid medicine comprises the following components: the mass fraction is 0.45 percent of sodium chloride, 4mg/ml betamethasone sodium phosphate and glucan, and the solvent is water. (experiments set different molecular weight and concentration of dextran, including molecular weight of 10000, mass fractions of 2%, 5%, 10% of T10-2%, T10-5%, T10-10%, molecular weight of 40000, mass fractions of 2%, 5%, 10% of T40-2%, T40-5%, T40-10%, molecular weight of 70000, mass fractions of 0.5%, 1%, 2% of T70-0.5%, T70-1%, T70-2%)
(3) Adding 100 μ l of hypertonic solution into the system in (2), mixing gently, standing and incubating at 37 deg.C for 30min, and mixing gently with shaking every 5 min. Wherein the hypertonic solution comprises the following components: the mass fraction is 4.5% of sodium chloride, 100mM of glucose and 100mM of sodium pyruvate, and the solvent is water.
(4) Centrifuging at 4 deg.C and 2000r/min for 4min, discarding supernatant, washing with 1ml pre-cooled physiological saline for 3 times, and removing dextran and free drug to obtain dextran-protected drug-loaded erythrocyte.
Blank control group:
the control drug-loaded red blood cells BSP-RBC were prepared according to the methods described in (1) to (4) above, with the only difference that the hypotonic solution contained no dextran.
Example 2 examination of the Effect of different molecular weights and different concentrations of dextran on the drug loading of drug-loaded erythrocytes
Collecting the supernatant of each step in example 1 and 4, sucking 200 μ l, adding 5 times volume of 1ml methanol, mixing, precipitating a little dextran and hemoglobin, filtering with 0.22 μm disposable filter membrane, introducing sample by high performance liquid chromatograph, and measuring the content of betamethasone sodium phosphate in the supernatant. The drug loading amount is the total amount of betamethasone sodium phosphate added-the content of betamethasone sodium phosphate in the supernatant.
Wherein the determination conditions of the high performance liquid chromatograph are as follows:
a chromatographic column: agilent
XB-C18(5 μm, 4.6X 250 nm); a detector: an Agilent technologies uv detector; mobile phase: methanol-0.05 mol/l potassium dihydrogen phosphate (1: 1); flow rate: 1 ml/min; column temperature: 40 ℃; detection wavelength: 254 nm.
The results in FIG. 1 show that the addition of dextran can increase the drug loading of erythrocytes. Compared with drug-loaded red blood cells BSP-RBC prepared without glucan, the drug-loaded red blood cells obtained by adding T10-5%, T10-10%, T40-5% and T40-10% are remarkably improved in drug-loaded rate. Represents p < 0.05, and has significant difference.
Example 3 examination of the Effect of different molecular weights and concentrations of dextran on osmotic fragility of drug-loaded erythrocytes
A series of sodium chloride solutions of different osmotic pressures were prepared with 1% NaCl and ultra pure water as shown in Table 1 below. 10 parts of the drug-loaded red blood cells prepared in the experimental group of example 1 were added with 4 times volume of sodium chloride solution of different osmotic pressures, and left to stand at 4 ℃ for 2 hours, and centrifuged at 4 ℃ and 2000r/min for 5 minutes, 100. mu.l of the supernatant was added to a 96-well plate, 3 parallel wells were provided for each sample, and the absorbance at 540nm of each sample well was measured with a microplate reader. The hemolysis rate was calculated according to the following formula, and the osmotic pressure was plotted as abscissa and the hemolysis rate was plotted as ordinate to draw an osmotic fragility curve. The penetration fragility determination method of the natural red blood cells and the blank control group BSP-RBC drug-loaded red blood cells is the same in 3 parallel tests.
TABLE 1
The results in fig. 2 show that the addition of dextran can enhance the ability of the drug-loaded erythrocytes to resist osmotic pressure, i.e., enhance the toughness of the drug-loaded erythrocytes. The improvement effect of T40-10% is especially obvious, and the penetration fragility of the drug-loaded red blood cells prepared by adding the glucan is almost the same as that of natural red blood cells.
Example 4 examination of dextran at different molecular weights and different concentrations for drug-loaded erythrocyte Na+/K+Influence of ATPase Activity
20 mul of the drug-loaded red blood cells prepared in the experiment set in the example 1 are added with 180 mul of ultrapure water for cracking and are kept stand for 30min to obtain transparent cracking liquid. Diluting 20 μ l of transparent lysate with ultrapure water 40 times until the lysate is colorless or transparent, measuring protein concentration with BCA protein concentration quantification kit, and measuring Na in the transparent lysate according to the procedure of ultramicro ATP enzyme kit+/K+-the activity of an ATPase. Finally, the activity of Na +/K + -ATP enzyme on the BSP-RBC membrane is converted according to the corresponding proportion. Natural red blood cells, blank control group BSP-RBC drug-loaded red blood cells Na+/K+The determination of ATPase activity is carried out in the same manner.
FIG. 3 shows that the addition of dextran can protect the drug-loaded erythrocyte Na+/K+-the activity of an ATPase. Wherein the protective effects of T40-5%, T40-10%, T70-1% and T70-2% are particularly remarkable.
Example 5 examination of the Effect of dextran on Phosphatidylserine eversion on membranes of drug-loaded erythrocyte
And (3) taking 10 mu l of the experimental group T40-10% -RBC group drug-loaded red blood cells in the example 1 to a flow tube, adding 490 mu l of 1 XPBS for dilution, taking 10 mu l of the diluted drug-loaded red blood cell suspension, adding 100 mu l of 1 XPinding buffer, fully mixing uniformly, adding 5 mu l of Annexin-V-FITC, and incubating for 10-15min at room temperature in a dark place. Add 400. mu.l of 1 XBinding buffer, mix well and test the sample. The experiment was set up with a Negative Control group (Negative Control), a natural red blood cell group (NRBC), a blank Control group carrying red blood cells (BSP-RBC), and a T40-10% -RBC group. Wherein, the negative control group is directly added with 500 mul of 1 × binding buffer without adding 5 mul of Annexin-V-FITC; the processing steps of the natural red blood cell group and the blank control group BSP-RBC drug-loaded red blood cells are the same as those of the T40-10% -RBC group.
The results in FIGS. 4A, 4B, 4C, and 4D show that Negative Control, NRBC, BSP-RBC, and T40-10% -RBC are distributed in FSC-SSC substantially uniformly, indicating that the morphology of the drug-loaded red blood cells is not affected by the addition of dextran and is substantially consistent with the morphology of the native red blood cells. The results in FIG. 4E show that FITC is the strongest in the BSP-RBC group, FITC positive rate is 26%, i.e., phosphatidylserine eversion rate is 26%, while FITC is significantly shifted to the left in the T40-10% -RBC group, FITC positive rate is reduced to 7.9%, i.e., phosphatidylserine eversion rate is 7.9%. Compared with the prior art, the p value is less than 0.05, and the obvious difference is shown. The addition of the T40-10% glucan can reduce the eversion of phosphatidylserine on an erythrocyte membrane and has good protection effect on erythrocytes.
Example 6 examination of the Effect of dextran on the morphology of drug-loaded erythrocytes
50 μ L of the T40-10% -RBC drug-loaded red blood cells of the experimental group in example 1 were added into an EP tube containing 1mL of 2.5% glutaraldehyde, mixed by gentle shaking, and fixed for 30min, during which the EP tube was gently shaken every 5min to prevent a large amount of red blood cells from precipitating at the bottom. The fixed red blood cell suspension is centrifuged at 2000r/min for 4min, the supernatant is removed, and the excess fixative is removed by washing 1-2 times with 1 XPBS buffer. The washed erythrocyte sediment is added into an EP tube filled with 1mL of after-fixing solution (0.4% potassium permanganate, 0.6% potassium dichromate), gently shaken and uniformly mixed, and suspended and fixed for 5 min. Centrifuging the fixed erythrocyte suspension for 4min at the rotating speed of 2000r/min, removing the supernatant, and washing with ultrapure water for 1-2 times to remove the redundant after-fixing solution. Then, the red blood cells are subjected to gradient dehydration by using ethanol dehydrating agents (30%, 50%, 70%, 80%, 85%, 90%, 95%, 100%) with different prepared concentrations. Suspending and dehydrating ethanol with each concentration for 5min, centrifuging at 2000r/min for 4min, removing supernatant, and dehydrating ethanol with the next concentration, wherein 100% ethanol is dehydrated twice. And finally, keeping 1/3 supernatant of the centrifuged cells, blowing and beating uniformly, dropping on a filter paper wafer, putting into a vacuum drying oven, and taking out the sample after 10 min. Dipping a small amount of sample powder on a conductive adhesive, plating gold, and observing by a scanning electron microscope. The treatment method of the natural red blood cells and the blank control group BSP-RBC drug-loaded red blood cells is the same.
The results in fig. 5 show that the addition of dextran does not affect the morphology of the drug-loaded red blood cells and is substantially consistent with the morphology of natural red blood cells, all in the shape of a biconcave round cake.
Example 7
The invention also refers to the experimental group of the embodiment 1, the drug-loaded red blood cells are prepared by other methods, and the drug-loaded quantity, the osmotic fragility and the red blood cells Na are treated+/K+Characterization of ATPase activity, eversion of phosphatidylserine on the membrane, morphology.
As a result: the drug-loaded erythrocytes prepared by the methods 1, 2 and 3 have the following characteristics: the addition of glucan improves the drug loading of erythrocytes, and the drug loading of erythrocytes prepared by adding T10-5%, T10-10%, T40-5% and T40-10% is remarkably improved (p is less than 0.05). The addition of the glucan enhances the osmotic pressure resistance of the drug-loaded red blood cells, wherein the T40-10% improvement effect is particularly obvious, and the osmotic fragility of the drug-loaded red blood cells prepared by adding the glucan is almost the same as that of all natural red blood cells. The addition of glucan protects the loaded red blood cells Na+/K+ATPase activity, with T40-5%, T40-10%, T70-1% and T70-2% being particularly significant. The addition of T40-10% dextran reduces the eversion of phosphatidylserine on erythrocyte membrane, has good protection effect on erythrocyte, and does not affect the shape of drug-loaded erythrocyte, which is basically the same as the shape of natural erythrocyte and is round cake-shaped with two concave sides.
While the invention has been described with respect to a preferred embodiment, it will be understood by those skilled in the art that the foregoing and other changes, omissions and deviations in the form and detail thereof may be made without departing from the scope of this invention. Those skilled in the art can make various changes, modifications and equivalent arrangements, which are equivalent to the embodiments of the present invention, without departing from the spirit and scope of the present invention, and which may be made by utilizing the techniques disclosed above; meanwhile, any changes, modifications and variations of the above-described embodiments, which are equivalent to those of the technical spirit of the present invention, are within the scope of the technical solution of the present invention.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811559344.0A CN109730975B (en) | 2018-12-19 | 2018-12-19 | A method for maintaining the biological activity of drug-loaded erythrocytes while increasing their drug-loading capacity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811559344.0A CN109730975B (en) | 2018-12-19 | 2018-12-19 | A method for maintaining the biological activity of drug-loaded erythrocytes while increasing their drug-loading capacity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109730975A CN109730975A (en) | 2019-05-10 |
| CN109730975B true CN109730975B (en) | 2021-07-09 |
Family
ID=66360708
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811559344.0A Active CN109730975B (en) | 2018-12-19 | 2018-12-19 | A method for maintaining the biological activity of drug-loaded erythrocytes while increasing their drug-loading capacity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109730975B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113318088A (en) * | 2021-06-16 | 2021-08-31 | 上海交通大学 | Compound medicine and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0824213A2 (en) * | 1996-08-15 | 1998-02-18 | Tsukasa Matsumoto | Method for fractionating red blood cells and antibacterial materials or bacterial proliferation inhibitors produced thereby |
| WO2008003524A2 (en) * | 2006-07-03 | 2008-01-10 | Università Degli Studi Di Urbino 'carlo Bo' | Delivery of contrasting agents for magnetic resonance imaging |
| CN106668840A (en) * | 2017-02-10 | 2017-05-17 | 南通大学 | Insulin controlled-release drug, and preparation method and application thereof |
| CN107802646A (en) * | 2016-09-05 | 2018-03-16 | 湖北盛齐安生物科技股份有限公司 | A kind of anti-tumor medicine |
| CN108113977A (en) * | 2018-01-25 | 2018-06-05 | 上海交通大学 | A kind of gelatin of erythrocyte membrane encapsulating carries the preparation method and applications of Berberine hydrochloride nanoparticle |
-
2018
- 2018-12-19 CN CN201811559344.0A patent/CN109730975B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0824213A2 (en) * | 1996-08-15 | 1998-02-18 | Tsukasa Matsumoto | Method for fractionating red blood cells and antibacterial materials or bacterial proliferation inhibitors produced thereby |
| WO2008003524A2 (en) * | 2006-07-03 | 2008-01-10 | Università Degli Studi Di Urbino 'carlo Bo' | Delivery of contrasting agents for magnetic resonance imaging |
| CN107802646A (en) * | 2016-09-05 | 2018-03-16 | 湖北盛齐安生物科技股份有限公司 | A kind of anti-tumor medicine |
| CN106668840A (en) * | 2017-02-10 | 2017-05-17 | 南通大学 | Insulin controlled-release drug, and preparation method and application thereof |
| CN108113977A (en) * | 2018-01-25 | 2018-06-05 | 上海交通大学 | A kind of gelatin of erythrocyte membrane encapsulating carries the preparation method and applications of Berberine hydrochloride nanoparticle |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109730975A (en) | 2019-05-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2023503711A (en) | A Lectin-Polymer Carrier Coupling Complex Used to Isolate Glycosylated Exosomes from Clinical Samples | |
| CS241478B2 (en) | Carrier system's particles and method of their production | |
| CN107266563A (en) | A kind of preparation method of hemoglobin quality-control product | |
| WO2007089665A2 (en) | Storage-stable cellular whole blood composition containing elevated amounts of d-dimer | |
| CN109730975B (en) | A method for maintaining the biological activity of drug-loaded erythrocytes while increasing their drug-loading capacity | |
| SU1767433A1 (en) | Method of determining insulin resistance of immunogenesis in patients with type 1 diabetes mellitus | |
| CN111513059A (en) | Red blood cell preservation solution and its application | |
| CN114414687A (en) | Binding buffer solution for lactoferrin extraction, preparation method and application thereof, and bovine lactoferrin detection method | |
| CN113509486B (en) | Application of CCA-CD co-assembly in preparation of macromolecular toxin detoxification drug | |
| IE44448B1 (en) | Gonococcal pili processes for the preparation thereof and the use thereof | |
| CN1683522A (en) | Whole blood quality control substance as cell bio-activity protector and its preparing method | |
| CN109453137A (en) | A kind of red blood cell living carries the sustained release preparation and the preparation method and application thereof of betamethasone sodium phosphate | |
| CN104535773B (en) | A kind of dust mite specific IgG4 subclass antibodies calibration object and preparation method thereof | |
| CN113318088A (en) | Compound medicine and preparation method and application thereof | |
| CN110812339A (en) | Red blood cell-encapsulated berberine hydrochloride composite system and preparation method and use thereof | |
| CN111471100B (en) | Novel erythrocyte membrane monoclonal antibody production method and processing device thereof | |
| CN107649102A (en) | A kind of preparation method of compound doughnut polymeric adsorbent | |
| CN111537326A (en) | Method for preparing freeze-dried blood platelet and application thereof | |
| CN106754692A (en) | A kind of erythrocyte membrane and its preparation method and application | |
| Blakeslee et al. | The J substance of cattle | |
| CN115399314B (en) | A kind of human ABO blood type anti-typing erythrocyte that can be freeze-dried and preserved and its preservation method | |
| CN117233388B (en) | Kit for detecting treponema pallidum antibodies in urine and preparation method thereof | |
| Nash et al. | Influence of parenteral nutrition on blood rheology and platelet aggregation in vitro | |
| CN117705541B (en) | Quality control product for detecting PD-1 of blood sample and preparation and quality control method thereof | |
| CN117654450B (en) | A material for adsorbing lipoprotein in blood and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |