CN100398151C - Ginsenoside nano microparticle and its preparing method and use - Google Patents
Ginsenoside nano microparticle and its preparing method and use Download PDFInfo
- Publication number
- CN100398151C CN100398151C CNB2005100491061A CN200510049106A CN100398151C CN 100398151 C CN100398151 C CN 100398151C CN B2005100491061 A CNB2005100491061 A CN B2005100491061A CN 200510049106 A CN200510049106 A CN 200510049106A CN 100398151 C CN100398151 C CN 100398151C
- Authority
- CN
- China
- Prior art keywords
- ginsenoside
- cholesterol
- lecithin
- solution
- centrifugal
- 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.)
- Expired - Fee Related
Links
Images
Abstract
The present invention discloses a ginsenoside nanometer particle and a preparation method and usage thereof. Ginsenoside, cholesterol and phosphatidyl choline are mixed according to the weight ratio of 20:1:1 and are assisted to be dissolved by a nonionic surface active agent, the ginsenoside, the cholesterol and the phosphatidyl choline interact to automatically form the nanometer particle, and the nonionic surface active agent is eliminated by a dialysis way or an ultrafiltration method. The formed ginsenoside nanometer particle (Ginsome) is observed to be a spherical micro particle of which the diameter is from 60 to 80 nm under an electron microscope, the Ginsome has the functions of a vaccine immunological adjuvant and the humoral immunity and cellular immunity functions of vaccine induction enhancement, the immunogenicity of antigen can be obviously increased only by 10 micrograms, and the ginsenoside nanometer particle has wide application in vaccine manufacture.
Description
Technical field
The present invention relates to a kind of ginsenoside nano microparticle (Ginsome) and its production and use.
Technical background
Adjuvant is the material that a class can strengthen vaccine-induced immunoreation intensity or change immunoreation type.After since eighties of last century Glenny twenties discovery aluminium hydroxide aluminium glue has adjuvant, the aluminium hydroxide aluminium glue is present unique vaccine for man adjuvant through approval, has been widely used in the production of people's epizootic disease Seedling.But aluminium hydroxide aluminium glue adjuvant has many weak points
(1.2.3.4), for example, 1) and it can promote IgE antibody to produce, and induces body to produce anaphylaxis easily; 2) injection back local irritation is bigger; 3) vaccine of multiple injection aluminium hydroxide aluminium glue can cause aluminum salt in localized accumulated, causes local muscle disease; 4) the aluminium hydroxide aluminium glue mainly promotes the reaction of Th2 para-immunity, and reaction promotes little to the Th1 para-immunity.The reaction of Th1 para-immunity mainly is at endotrophic pathogenic microorganism; The reaction of Th2 para-immunity mainly is at ectoparasitic pathogenic microorganism of cell and parasite.So the aluminium hydroxide aluminium glue is mainly used in the production of bacillary vaccine, and has little significance for the adjuvant of some viral vaccines.
Traditional vaccine is generally made with whole cell or totivirus, contains multiple antigenic component, can produce the small part of having only of protective immunological reaction in the middle of them, not only remaining antigen there is no need, also can cause certain side reaction.Along with going deep into of amynologic basis theoretical research, and the development of antigen purification technique and technique for gene engineering, many subunit antigen and polypeptide antigen have appearred, and the antigen that has can synthetic.Compare with traditional bacteria vaccine, these antigens have the advantage that molecular weight is little, purity is high, safety is good.But the relevant vaccine of succeeding in developing up to now, is but very few for number.One of its reason is an immunogenicity of antigens variation behind the purification, is difficult to induce body to produce enough immunoreation.
Therefore, research had not only promoted the Th1 class but also had promoted the novel adjuvant of Th2 para-immunity reaction or improved the immune effect of subunit vaccine significant.We in the past studies show that the ginsenoside is the immunological adjuvant of a safety.It not only can improve the reaction of Th1 para-immunity, also can improve the reaction of Th2 para-immunity
(5.6.7.8. 9)Relevant achievement has obtained national inventing patent
(10)The ginsenoside can be from the root of Chinese herbal medicine Radix Ginseng Panaxginseng C.A.Mey, the root of Radix Panacis Quinquefolii Panax quinquefolium L., the root of Radix Notoginseng Panaxnotoginseng (Burk.) F.H.Chen, perhaps extracting in the herb of Herb Gynostemmae Pentaphylli Gynostemma pentaphyllum (Thunb.) Mak., is the chemical compound that a class has the triterpenoid saponin chemical constitution
(11)The extracting method that the ginsenoside uses always is
(12): extract with ethanol or methanol solvate, reclaim solvent, residue is water-soluble, the filtering insoluble matter, aqueous solution is made two-phase extraction with lipotropy organic solvents such as petroleum ether, benzene or ether, to remove lipotropy impurity.Use water saturated butanols extraction then instead, Saponin is shifted in butanols.Collect butanols, evaporated under reduced pressure, rough ginsenoside.Rough ginsenoside is dissolved in small amount of methanol or the ethanol, dropwise adds acetone then, mix homogeneously is promptly separated out the ginsenoside.So handle for several times, can obtain refining ginsenoside.So the ginsenoside who obtains needs more heavy dose of competence exertion adjuvant effect, and is directly higher as the immunological adjuvant relative cost of animal vaccine.
List of references
1.Lindblad，L.B.&Sparck J.V.(1987)Basic concepts in the applicationof immunological adjuvants.Scand.J.Lab.Anim.Sci.14：1-13
2.Bomford，R.(1994)Adjuvants.In：Dale，M.M.et al.(Ed)Textbookof Immunopharmacology，3
rd ed，Blackwell Scientific Publications，Oxford.pp.335-339
3.Ott，G.Nest，G.V.&Burke，R.L.(1994)The use of Muramyl peptidesas vaccine adjuvants.In：International Business Communications-Newadvances in vaccine technologies and applications February 23-25，1994，pp.89-114
4.Malakoff，D.Public Health.Aluminum is put on trial as a vaccinebooster.Science 2000，288，1323-1324
5.HU，S.CONCHA，C，COORAY，R.HOLMBERG，O.Ginseng-enhanced oxidative andphagocytic activities of polymorphonuclear leucocytes from bovineperipheral blood and stripping milk.Vet.Res.1995，26，155-161
6.CONCHA，C.HU，S.HOLMBERG，O.The proliferative responses of cowstripping milk and blood lymphocytes to pokeweed mitogen and ginsengin vitro.Vet.Res.1996，27，107-115
7.HU，S.CONCHA，C.LIN，F.PERSSON WALLER，K.Adjuvant effect of ginsengextracts on the immune responses to immunization againstStaphylococcus aureus in dairy cattle.Vet.Immunol.Immunopathol.2003，91，29-37
8.RIVERA，E.DAGGFELDT，A.HU，S.Ginseng extract in aluminium hydroxideadjuvanted vaccines improves the antibody response of pigs to porcineparvovirus and Erysipelothrix rhusiopathiae.Vet.Immunol.
Immuopathol.2003，91，19-27
9.Rivera，E.Hu，S.Concha，C.Ginseng and aluminium hydroxide actsynergistically as vaccine adjuvants.Vaccine 2003，21，1149-1157
10. the vaccine adjuvant purposes (patent No.: ZL02110578.2) of Radix Ginseng total saponins and saponin monomer Rb1;
11. the little younger sister of Song, Tang Zhishu (chief editor), Chinese herbal medicine chemical constituent extraction separation and preparation (the 1st edition), People's Health Publisher, Beijing,, the 33rd page, the 51st page, the 206th page, the 394th page in 2004.
12. Wang Xian pattern (chief editor), Natural Medicine Chemistry, the 1st edition, the People's Health Publisher publishes, Beijing,, the 549th page in 1988.
Summary of the invention
The purpose of this invention is to provide a kind of ginsenoside nano microparticle (Ginsome) and its production and use.
Ginsenoside nano microparticle (Ginsome) is that the diameter of a kind of ginsenoside of containing, cholesterol and lecithin is the spheroidal particle of 60～80nm.
The step of the preparation method of ginsenoside nano microparticle (Ginsome) is as follows:
1) preparation of ginsenoside's solution: the ginsenoside is dissolved in distilled water, is made into 50～100mg/ml solution, filtration sterilization, standby;
2) preparation of cholesterol/lecithin soln: with nonionic surfactant, Mega-10 for example, available Sigma company's product or octyl group glycoside octylglucoside, be configured to 20～25% aqueous solution, then lecithin and cholesterol are dissolved in the nonionic surfactant solution, make it contain lecithin and each 10～15 mg/ml of cholesterol, filtration sterilization, standby;
3) above-mentioned ginsenoside's solution and cholesterol/lecithin soln are mixed, make the ginsenoside, cholesterol and lecithin three's weight ratio is 20: 1: 1, add phosphate buffer, pH7～7.4 make mixed solution contain ginsenoside 20's mg/ml, cholesterol 1 mg/ml and lecithin 1 mg/ml, mixing is put 25～50 ℃ of waters bath with thermostatic control 0.5～2 hour;
4) mixing material being transferred to molecular cut off is in 12,000～14,000 the bag filter, then bag filter is put into phosphate buffer, pH7.4, middle dialysis, the dialysis temperature is 20～40 ℃, changed phosphate buffer one time in per 6 hours, dialysed continuously 24～72 hours, the liquid in the bag filter is transferred in the centrifuge tube, centrifugal 10～30 minutes, centrifugal speed is 400～500g, abandons precipitation, gets final product;
Perhaps, it is in 10,000～100,000 the centrifugal ultrafiltration pipe that mixing material is transferred to molecular cut off, the centrifugal ultrafiltration pipe can adopt Millipore company product (
Centrifugal Filter Units, www.millipore.com/puredna) centrifugal, centrifugal speed is 500～14,000g adds normal saline and continues centrifuge washing, so repeats 2～6 times.Add normal saline again, mixing gets final product.
Ginsenoside nano microparticle is used for the immunological adjuvant of vaccine.
Advantage of the present invention
1) ginsenoside nano microparticle Ginsome can be used as the immunological adjuvant of subunit vaccine;
2) ginsenoside makes after the nanoparticle Ginsome, the ginsenoside strengthens the adjuvant effect of subunit vaccine, significantly reduce and reach the needed ginsenoside's dosage of performance adjuvant effect, therefore reduced cost, Ginsome can be used as the novel adjuvant of developing subunit vaccine from now on;
3) Ginsome can promote Th1 class and the reaction of Th2 para-immunity, therefore, can be used as the immunological adjuvant of multiple vaccine, brings into play better immune effect;
4) Ginsome can stored frozen, still keeps the Nano microsphere form constant after thawing, and can preserve for a long time.
Description of drawings
Fig. 1 is Ginsome electron micrograph (phosphotungstic acid dyeing, 30000 times of amplifications).Ginsenoside nano microparticle (Ginsome) is that the diameter of a kind of ginsenoside of containing, cholesterol and lecithin is the spheroidal particle of 60～80nm;
Fig. 2 is back 3 the anti-OVA antibody test of the serum result schematic diagrams of two immunity.
The specific embodiment
The preparation method one of ginsenoside nano microparticle (Ginsome)
1. the preparation of ginsenoside's solution: the ginsenoside is dissolved in distilled water, is made into 50～100mg/ml solution, filtration sterilization, standby.
2. the preparation of cholesterol/lecithin soln: with nonionic surfactant Mega-10 (Sigma company product); be configured to 20% aqueous solution; then lecithin (Chinese workers factory of Chinese East China Normal University product) and cholesterol (Shanghai chemical reagents corporation product) are dissolved in Mega-10 (the Sigma company product) solution; make it contain lecithin and each 10～15 mg/ml of cholesterol; filtration sterilization, standby.
3. above-mentioned ginsenoside's solution and cholesterol/lecithin soln are mixed, make the ginsenoside, cholesterol and lecithin three's weight ratio is 20: 1: 1, add phosphate buffer, pH7.4 makes mixed solution contain ginsenoside 20's mg/ml, cholesterol 1 mg/ml and lecithin 1 mg/ml, mixing is put 37 ℃ of waters bath with thermostatic control 1 hour.
4. mixing material is transferred to molecular cut off and is in 12,000～14,000 the bag filter, then bag filter is put into phosphate buffer, pH7.4, middle dialysis, the dialysis temperature is 37 ℃, changed phosphate buffer one time in per 6 hours, dialysed continuously 48 hours, the liquid in the bag filter is transferred in the centrifuge tube, centrifugal 20 minutes, centrifugal speed is 470g, abandons precipitation, gets final product.
The preparation method two of ginsenoside nano microparticle (Ginsome)
1. the preparation of ginsenoside's solution: the ginsenoside is dissolved in distilled water, is made into 50～100mg/ml solution, filtration sterilization, standby.
2. the preparation of cholesterol/lecithin soln: with nonionic surfactant Mega-10 (Amresco company product), be configured to 20% aqueous solution, then lecithin and cholesterol are dissolved in the Mega-10 solution, make it contain lecithin and each 10～15 mg/ml of cholesterol, filtration sterilization, standby.
3. above-mentioned ginsenoside's solution and cholesterol/lecithin soln are mixed, make the ginsenoside, cholesterol and lecithin three's weight ratio is 20: 1: 1, add phosphate buffer, pH7.4 makes mixed solution contain ginsenoside 20's mg/ml, cholesterol 1 mg/ml and lecithin 1 mg/ml, mixing is put 37 ℃ of waters bath with thermostatic control 1 hour.
4. mixing material is transferred to molecular cut off and is in 10,000～100,000 the centrifugal ultrafiltration pipe, the centrifugal ultrafiltration pipe be Millipore company product (
Centrifugal Filter Units, www.millipore.com/puredna) centrifugal, centrifugal speed is 8000g, adds normal saline and continues centrifuge washing, so repeats 5 times.Add normal saline again, mixing gets final product.
Ginsenoside nano microparticle (Ginsome) is to oralbumin (OVA) immunoadjuvant function of pattern subunit antigen
Material and method
1. the mixed liquor of pattern antigen and adjuvant: pattern antigen oralbumin (OVA, Shanghai uncle's biotechnology difficult to understand company product) is dissolved in normal saline.Then with OVA and ginsenoside (GS) or the mixing of ginsenoside nano microparticle (Ginsome) solution, make every milliliter and contain 50 μ g OVA, and every milliliter of solution that contains 50 μ g OVA and 250 μ g ginsenosides or Ginsome, shaking table shook in 24 hours, fully mixing is put refrigerator (4 ℃) then and is preserved standby.
2. laboratory animal and immunization method thereof: 35 of 8～10 female SPF Balb/c mices in age in week (Shanghai country laboratory rodent kind subcenter provides).Mice is divided into 7 groups at random, 5 every group.The oralbumin (OVA) of every mouse carotid back subcutaneous injection pattern subunit antigen or OVA and immunological adjuvant mixture are injected once after 28 days at interval again, and the per injection object is long-pending to be 0.2 milliliter, sees Table 1.
The grouping of table 1 laboratory animal reaches respectively organizes processing mode
3. immunity back is observed: observe the mice mental status after the per injection, behavior changes, claim mice body weight and record every day, record blood sampling situation and mice change.
4. blood specimen collection: after the immunity second time 1,2,3 weeks, every aseptic indoor capillary tube eye socket blood vessel blood sampling of mice, blood is put the 0.5mL plastic centrifuge tube, in 4 ℃ of refrigerator overnight, second day centrifugal, and centrifugal speed is 6000 rev/mins, centrifugal 20 minutes, with serum transfers to the packing of 0.2mL plastic tube ,-20 ℃ of preservations, standby.
5. antibody test: every hole adds 100 μ l coating buffer (the 0.05mol/L carbonate buffer solutions that contain 1.4mgOVA/ml on 96 hole ELISA Plate, pH9.6), put 4 ℃ hatch 24 hours after, with cleaning mixture (0.01mol/LPBS-0.05% tween 20, pH7.4) 3 times (every hole 200 μ l) of washing, each 3 minutes.Every hole adds 300 μ l confining liquids (0.01mol/L PBS-3% tween 20 solution), and incubated at room is after 1 hour, with 3 times (every hole 300 μ l) of cleaning mixture washing, each 3 minutes.Add serum to be checked and negative serum (dilution in 1: 100), every hole 100 μ l.Behind the incubated at room 2h, with 3 times (every hole 200 μ l) of cleaning mixture washing, each 3 minutes.The goat anti-mouse IgG antibody (Chemicon International, the Inc. product that add horseradish peroxidase-labeled; Dilution in 1: 500), 100 μ l/ holes.After the incubated at room 2 hours, with 3 times (every hole 200 μ l) of cleaning mixture washing, each 3 minutes.Add the colour developing of tetramethyl biphenyl diamidogen (TMB, SIGMA-ALDRICH company product) solution, 100 μ l/ holes, 37 ℃ hatch 30 minutes after, add 2M H
2SO
4Solution 50 μ l/ hole cessation reactions.Put microplate reader (Multiskan MK3) and measure OD
450(absorbance, OD) value.
Result and analysis
1. ginsenoside nano microparticle (Ginsome) subcutaneous injection is safety to Mus
After twice immunity, each organizes mice in all no abnormal variations in aspect such as appetite, the mental status, ability to acts.Body weight is weightening finish trend descending to some extent substantially after week after the 1st immunity; Two exempt from the no weight loss situation in back.Table 2 is the average weight of every group of 4 body weight weighing of mice.Each organizes mice body weight change there was no significant difference.
Average weight (the unit: g) of every group of mice of table 2
2. the anti-OVA antibody horizontal of immune back serum
Fig. 2 is back 3 the anti-OVA antibody test of the serum results of two immunity, illustrates: 1) two immunity detect for back 3 times, and the 2nd group antibody horizontal all is lower than the 1st group, but no difference of science of statistics, the ginsenoside is when dosage is 50 micrograms in expression, no adjuvant effect; 2) the 3rd, 4,5,6,7 group antibody horizontal is significantly higher than the 1st, 2 group, illustrates that ginsenoside nano microparticle (Ginsome) has adjuvant effect; 3) ginsenoside's dosage of the 2nd group is 50 micrograms, and the 3rd, the 4 group of contained ginsenoside's dosage of Ginsome is respectively 10,50 micrograms.People such as Rivera report (Rivera, E.Hu, S.Concha, C.Ginseng and aluminiumhydroxide act synergistically as vaccine adjuvants.Vaccine 2003,21,1149-1157), the ginsenoside does not have adjuvant effect when dosage is 160 micrograms, and adjuvant effect is arranged when 830 micrograms.This test is prepared into ginsenoside nano microparticle (Ginsome) with the ginsenoside, at the dosage range that contains ginsenoside's 10～1000 micrograms remarkable adjuvant effect is arranged all, therefore, the ginsenoside makes after the nanoparticle (Ginsome), adjuvant effect strengthens, and ginsenoside's consumption reduces.
Embodiment 4
Ginsenoside nano microparticle (Ginsome) is to the adjuvant effect of cracking influenza virus vaccine
Experimental technique
1. the mixed liquor of antigen and adjuvant: with cracking influenza virus (H
3N
2Strain, Tianyuan biotech firm in Hangzhou provides) with the normal saline dilution or and Ginsome or and aluminium hydroxide mix, be mixed with and contain following material in per 0.2 milliliters of liquid: 1) cracking influenza virus, 50 HAUs (HAU); 2) cracking influenza virus, 100HAU; 3) cracking influenza virus, 500HAU; 4) cracking influenza virus, 100HAU+Ginsome (containing ginsenoside's 10 micrograms); 5) cracking influenza virus, 100HAU+50 microgram aluminium hydroxide.Fully mixing is put refrigerator (4 ℃) then and is preserved standby.
2. laboratory animal and immunization method thereof: 30 of the female SPF Balb/c mices of body weight 18～20 grams (Shanghai country laboratory rodent kind subcenter provides).Mice is divided into 5 groups at random, 6 every group.Every above-mentioned 5 kinds of liquid of mice abdomen thigh portion subcutaneous injection are injected once after 21 days at interval again, and the per injection object is long-pending to be 0.2 milliliter, sees Table 1.
Blood specimen collection and blood clotting suppress to tire detection
4 weeks after the immunity second time, every aseptic indoor capillary tube eye socket blood vessel blood sampling of mice, blood is put the 0.5mL plastic centrifuge tube, in 4 ℃ of refrigerator overnight, second day centrifugal, and centrifugal speed is 6000 rev/mins, centrifugal 20 minutes, with serum transfers to the packing of 0.2mL plastic tube ,-20 ℃ of preservations, standby.Adopt blood clotting inhibition method to detect the mice serum blood clotting and suppress to tire (HI).
Result and analysis
Two exempt from the back blood clotting testing result that suppresses to tire shows that increase antigen dose merely, the HI antibody titer changes little; Conventional adjuvant aluminium hydroxide aluminium glue and matched group relatively do not have significant difference; Add after the ginsenoside nano microparticle Ginsome in the vaccine, HI tires and is significantly higher than matched group (table 3).
Behind table 3 injection various dose and the influenza vaccines that contain different adjuvants, mice serum HI tire (n=6)
| Inactivating influenza virus dosage (HAU, per injection dosage) | Adjuvant | Blood clotting suppresses to tire (HI, geometrical mean) |
| 50 | No |
1∶256 |
| 100 | No adjuvant | 1∶256 |
| 500 | No adjuvant | 1∶469 |
| 100 | Ginsome (containing 10 microgram ginsenosides) | 1∶683 |
| 100 | Aluminium hydroxide aluminium glue (50 microgram) | 1∶329 |
Claims (3)
1. a ginsenoside nano microparticle is characterized in that, it is that the diameter of a kind of ginsenoside of containing, cholesterol, lecithin is the spheroidal particle of 60～80nm, the ginsenoside, and cholesterol and lecithin three's weight ratio is 20: 1: 1.
2. the preparation method of a ginsenoside nano microparticle is characterized in that, the step of method is as follows
1) preparation of ginsenoside's solution: the ginsenoside is dissolved in distilled water, is made into 50～100mg/ml solution, filtration sterilization, standby;
2) preparation of cholesterol/lecithin soln: with nonionic surfactant Mega-10 or octyl group glycoside, be configured to 20～25% aqueous solution, then lecithin and cholesterol are dissolved in the nonionic surfactant solution, make it contain lecithin and each 10～15 mg/ml of cholesterol, filtration sterilization, standby;
3) above-mentioned ginsenoside's solution and cholesterol/lecithin soln are mixed, make the ginsenoside, cholesterol and lecithin three's weight ratio is 20: 1: 1, add phosphate buffer, pH7～7.4 make mixed solution contain ginsenoside 20's mg/ml, cholesterol 1 mg/ml and lecithin 1 mg/ml, mixing is put 25～50 ℃ of waters bath with thermostatic control 0.5～2 hour;
4) mixing material being transferred to molecular cut off is in 12,000～14,000 the bag filter, then bag filter is put into phosphate buffer, pH7.4, middle dialysis, the dialysis temperature is 20～40 ℃, changed phosphate buffer one time in per 6 hours, dialysed continuously 24～72 hours, the liquid in the bag filter is transferred in the centrifuge tube, centrifugal 10～30 minutes, centrifugal speed is 400～500g, abandons precipitation, gets final product;
Perhaps,
It is that centrifugal, centrifugal speed is 500～14 in 10,000～100,000 the centrifugal ultrafiltration pipe that mixing material is transferred to molecular cut off, and 000g adds normal saline and continues centrifuge washing, so repeats 2～6 times, adds normal saline again, and mixing gets final product.
3. the preparation method of a kind of ginsenoside nano microparticle according to claim 2, it is characterized in that, described ginsenoside is the root of Radix Ginseng, the root of Radix Panacis Quinquefolii, the root of Radix Notoginseng or the herb extract of Herb Gynostemmae Pentaphylli, and it is the chemical compound with triterpenoid saponin chemical constitution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100491061A CN100398151C (en) | 2005-02-18 | 2005-02-18 | Ginsenoside nano microparticle and its preparing method and use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100491061A CN100398151C (en) | 2005-02-18 | 2005-02-18 | Ginsenoside nano microparticle and its preparing method and use |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1683012A CN1683012A (en) | 2005-10-19 |
| CN100398151C true CN100398151C (en) | 2008-07-02 |
Family
ID=35262630
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100491061A Expired - Fee Related CN100398151C (en) | 2005-02-18 | 2005-02-18 | Ginsenoside nano microparticle and its preparing method and use |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100398151C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361971B (en) * | 2008-10-10 | 2011-01-26 | 福建省农业科学院生物技术研究所 | Preparation technique of Vibrio vulnificus whole-cell immunity stimulated compound vaccine and use thereof in aquatic animal |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101428145B (en) * | 2007-11-05 | 2013-01-02 | 北京生泰尔生物科技有限公司 | Novel vaccine adjuvant |
| CN113750229A (en) * | 2021-10-18 | 2021-12-07 | 浙江省农业科学院 | Nano granules of saponin of ginseng stem and leaf, preparation method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992021331A1 (en) * | 1991-05-31 | 1992-12-10 | Iscovent Ab | Pharmaceutical carrier |
| CN1141143C (en) * | 2002-01-16 | 2004-03-10 | 浙江大学 | Ginseng total saponin or monomer saponin RbI vaccine immunological adjuvant application |
-
2005
- 2005-02-18 CN CNB2005100491061A patent/CN100398151C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992021331A1 (en) * | 1991-05-31 | 1992-12-10 | Iscovent Ab | Pharmaceutical carrier |
| CN1141143C (en) * | 2002-01-16 | 2004-03-10 | 浙江大学 | Ginseng total saponin or monomer saponin RbI vaccine immunological adjuvant application |
Non-Patent Citations (4)
| Title |
|---|
| 中药纳米化的作用及存在的问题. 张晓静.江苏中医药，第24卷第3期. 2003 |
| 中药纳米化的作用及存在的问题. 张晓静.江苏中医药，第24卷第3期. 2003 * |
| 中药超微粉碎研究进展与思考. 胡学军等.世界科学技术-中药现代化，第4卷第5期. 2002 |
| 中药超微粉碎研究进展与思考. 胡学军等.世界科学技术-中药现代化，第4卷第5期. 2002 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361971B (en) * | 2008-10-10 | 2011-01-26 | 福建省农业科学院生物技术研究所 | Preparation technique of Vibrio vulnificus whole-cell immunity stimulated compound vaccine and use thereof in aquatic animal |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1683012A (en) | 2005-10-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| DE60121136T2 (en) | Proteasome-INFLUENZA VIRUS VACCINE COMPOSITION | |
| CN105903012B (en) | Pharmaceutical product comprising mite allergen extract and method for preparing the same | |
| JP5391282B2 (en) | Composition for promoting hematopoietic stem cell proliferation comprising fistula extract, reishi extract and mulberry extract | |
| Qiu et al. | Immunopotentiating effects of four Chinese herbal polysaccharides administered at vaccination in chickens | |
| AU2007305076A1 (en) | Immune modulators, preparations and compositions including immune modulators, tests for evaluating the activity of immune modulators and preparations and compositions including the same, and methods | |
| Li et al. | The selenylation modification of epimedium polysaccharide and isatis root polysaccharide and the immune-enhancing activity comparison of their modifiers | |
| CN101357883A (en) | Antrodia camphoratea pimelie kelone compound for treating autoimmune disease and medicine composition | |
| WO2020093510A1 (en) | Separation and purification method for polysaccharide in ganoderma lucidum spores | |
| CN100398151C (en) | Ginsenoside nano microparticle and its preparing method and use | |
| CN101670103A (en) | Compound panax ginseng total saponin and levamisole nano emulsion adjuvant and preparation method thereof | |
| CN105030562B (en) | A kind of antiallergy complex polysaccharide composition and its preparation method and application | |
| CN101869706A (en) | Compound astragalus polysaccharides and echinacea purpurea herb nanoemulsion adjuvant and preparation method thereof | |
| Zwickey et al. | The effect of Echinacea purpurea, Astragalus membranaceus and Glycyrrhiza glabra on CD25 expression in humans: a pilot study | |
| CN104352551B (en) | A kind of radix glycyrrhizae electromagnetic dissociation powder and its preparation method and application | |
| KR101787124B1 (en) | Arabinogalactan for enhancing the adaptive immune response | |
| WO2016141774A1 (en) | Uses of jilin ginseng oligopeptide in preparing food product or healthcare food product for improving and enhancing sexual function | |
| US6921543B2 (en) | Immunomodulatory preparation | |
| WO2002098437A1 (en) | Korean acanthopanax senticosus extract, protein extract, crude protein-polysaccharide which were extracted from korean acanthopanax senticosus, and immunoregulating compositions comprising the same and use thereof | |
| Yudhawan et al. | Immunomodulatory effect of Standardized Polysaccharide Fraction syrup from Noni fruit (Morinda citrifolia) on Cytokines level (IL-2 and IFN-γ) and Its Histological Evaluation in rats Vaccinated with Hepatitis-B | |
| CN104306392B (en) | Be cracked application of the Lasiosphaera fenzlii granulose in antioxidant is prepared | |
| CN109010325B (en) | Application of stilbene compounds derived from hemp leaf in preparing lipid-lowering medicines | |
| CN101380472B (en) | Use of total saponin from Panax notoginseng stalks and leaves in preparing immunoadjuvants | |
| Cohn | Quantitative distribution of phosphorus in chorioallantoic membrane as affected by infection with influenza virus. | |
| CN105169386B (en) | A kind of novel universal type matrix vaccines adjuvant and its preparation method and application | |
| CA2437690A1 (en) | Ganoderma lucidum spores for treatment of autoimmune diseases |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20080702 Termination date: 20110218 |