CN1211121C - Quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis B surface antigen, and prepn. method thereof - Google Patents
Quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis B surface antigen, and prepn. method thereof Download PDFInfo
- Publication number
- CN1211121C CN1211121C CNB018019250A CN01801925A CN1211121C CN 1211121 C CN1211121 C CN 1211121C CN B018019250 A CNB018019250 A CN B018019250A CN 01801925 A CN01801925 A CN 01801925A CN 1211121 C CN1211121 C CN 1211121C
- Authority
- CN
- China
- Prior art keywords
- combined vaccine
- dtwph
- toxoid
- concentration range
- hepatitis
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/29—Hepatitis virus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
- A61K39/29—Hepatitis virus
- A61K39/292—Serum hepatitis virus, hepatitis B virus, e.g. Australia antigen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/70—Multivalent vaccine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0016—Combination vaccines based on diphtheria-tetanus-pertussis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2730/00—Reverse transcribing DNA viruses
- C12N2730/00011—Details
- C12N2730/10011—Hepadnaviridae
- C12N2730/10111—Orthohepadnavirus, e.g. hepatitis B virus
- C12N2730/10134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Immunology (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Mycology (AREA)
- Communicable Diseases (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Oncology (AREA)
- Gastroenterology & Hepatology (AREA)
- Pulmonology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
A quadrivalent (DTwPH) combination vaccine comprising diphtheria toxoid, tetanus toxoid, whole cell pertussis, and HBsAg, and a method for preparing the same are provided. In the preparation of the DTwPH combination vaccine, diphtheria toxoid and tetanus toxoid are adsorbed onto aluminum phosphate (AlPO4) gel, and HBsAg is adsorbed onto aluminum hydroxide (Al(OH)3) gel. The DTwPH combination vaccine is adjusted to have a final pH of 6.5-7.5, and the concentrations of constituents also adjusted with the concentration of aluminum hydroxide gel in the range of 15-35 mu gAl/mL.
Description
Invention field
The present invention relates to a kind of quadrivalent combination vaccine (to call the DTwPH combined vaccine in the following text) that comprises diphtheria toxoid, tetanus toxoid, whole cell pertussis Seedling and hbs antigen and preparation method thereof.
In the expansion Immunization programme (EPI) of World Health Organization (WHO) (WHO), recommend to inoculate simultaneously the almost basic vaccine of the baby of whole world country use of diphtheria, tetanus and the conduct of pertussal DTP combined vaccine.In Southeast Asia, the most general hepatitis B infected in the Central Asia, Africa and Southeast Asia, South America, be considered to a kind of potential cause of hepatocarcinoma.Yet, hepatitis B infectedly also can prevent, so WHO actively develops whole world Hepatitis B virus vaccine inoculation activity by immunity.
Recently, have the DTP combined vaccine of different vaccination program and the trouble of Hepatitis B virus vaccine in order to eliminate independent inoculation, and be supplied to undeveloped country to increase its vaccination rate with the price that reduces these vaccines, now existing business-like combined vaccine product (DTaPH) (the European patent publication 0642355B1 that comprises diphtheria toxoid, tetanus toxoid, acellular pertussis and HBsAg, June 15,1998; Korean patent publication 1992-0009729, green cross company).
Background of invention
With the relevant active research of having carried out of preparation of DTwPH combined vaccine, solving the problem that runs in this combined vaccine medication preparation, HBsAg is immunogenic to prevent toxic increase simultaneously keeping.
The problem that takes place in the combined vaccine medication preparation comprises the adhesion to the glass storage bottle of the gathering of single vaccine solution and vaccine.Here, term is assembled " finger-type becomes slightly to shake the solid matter that can not make its suspension, and term " adhesions " refers to that DTwPH combined vaccine granule is attached to be preserved on the vial wall, in conjunction with very firm, slightly shake and can not make its separation.Gathering in the DTwPH combined vaccine is by especially causing as the gel aluminum hydroxide of adsorbent and the electrostatic attraction between the bordetella pertussis between each vaccine adsorbent solution and the bordetella pertussis.This adhesion is caused by secular reaction between DTwPH combined vaccine granule and the vial wall.Because DTwPH combined vaccine long term store is assembled and adhesion seriously reduces the stability of combined vaccine product.
The DTwPH combined vaccine is owing to adsorbent and the excessive problem with toxicity increase of albumen.In addition, because HBsAg and other antigen compatibility are poor, compare with Hepatitis B virus vaccine with the vaccine DPT combined vaccine that the while inoculating two kinds is independent, its immunogenicity reduces (WHO general headquarters, diph/tet-pertussis-hepatitis B tetravalent vaccine informal meeting, 1992).
Therefore, the present invention seeks to effectively to prevent above-mentioned " gathering " and " adhesion " phenomenon, a kind of new DTwPH combined vaccine and preparation method thereof also is provided, the immunogenicity that can keep HBsAg makes it and inoculates independent trivalent DTP combined vaccine and single antigen Hepatitis B virus vaccine simultaneously and have identical geometric average antibody titre (ab GMT) and potent antibodies production rate.
Detailed Description Of The Invention
The invention provides a kind of new tetravalence DTwPH combined vaccine, comprise diphtheria toxoid, tetanus toxoid, whole cell pertussis and HBsAg, wherein diphtheria toxoid and tetanus toxoid are adsorbed in Fosfalugel (Yamanouchi) (AlPO
4), HBsAg is adsorbed in gel aluminum hydroxide (Al (OH)
3).
The present invention also provides the preparation method of DTwPH combined vaccine, comprised (a) before preparing final DTwPH combined vaccine composition, regulating the Fosfalugel (Yamanouchi) solution of absorbed diphtheria toxoid, the Fosfalugel (Yamanouchi) solution of adsorbed tetanus toxoid, the gel aluminum hydroxide solution of absorption HBsAg and the pH scope of pertussis stock solution is 6.5-7.5; (b) diphtheria toxoid and the tetanus toxoid adsorbent solution that will adjust pH mixes; (c) mixture that will obtain in step (b) mixes with the HBsAg adsorbent solution of having adjusted pH and deactivation pertussis stock solution.
The present invention also provides the combined vaccine storage practice, comprises gelatin is added common combined vaccine or above-mentioned DTwPH combined vaccine, or uses the storage bottle of the vial of silicon coating as combined vaccine.The concentration range of preferred gelatin is 0.1-0.7w/v%, and is preferred, is 0.5-0.7w/v%.
Below structure of the present invention and effect will be described in more detail.
The diphtheria toxoid stock solution that the present invention uses is by cultivating diphtheria corynebacterium (Corynebacterium diphtheriae) Park Willium strain preparation.With purification behind the culture supernatant filtration, precipitation and the formalin detoxification that produce, obtain suitable diphtheria toxoid stock solution.Can use above-mentioned bacterial strain and method in addition to obtain suitable diphtheria toxoid stock solution.
The tetanus toxoid stock solution is by cultivating clostridium tetani (Clostridium tetani) Harvard strain preparation.With purification behind the culture supernatant filtration, precipitation and the formalin detoxification that produce, obtain suitable diphtheria toxoid stock solution.Can use above-mentioned bacterial strain and method in addition to obtain suitable diphtheria toxoid stock solution.
The pertussis stock solution is by cultivating bordetella pertussis (Bordetella pertussis) preparation.With the medium centrifugal that produces.The pertussis precipitation that obtains is suspended in the saline and by handling deactivation, obtains suitable deactivation pertussis stock solution.Can use above-mentioned bacterial strain and method in addition to obtain suitable pertussis stock solution.
HBsAg extracts by homogenate from the recombination yeast culture.The a series of separation method of HBsAg process from recombination yeast comprises ion chromatography, ultrafiltration, dialysis, ultracentrifugation, gel permeation chromatography etc., obtains suitable HBsAg stock solution.Can obtain suitable HBsAg by using above-mentioned method in addition, also can be used to HBsAg from blood plasma.
The suitable adsorbent that the present invention uses comprises aluminium hydroxide and Fosfalugel (Yamanouchi).Aluminium hydroxide and Fosfalugel (Yamanouchi) can be bought or prepare with known method.
The silicon coating bottle that is used among the present invention preserving combined vaccine was made vial in 10 minutes at 310 ℃ of heat dryings then by vial being immersed in 1: 40 diluent of 35% polymethyl siloxane (Dow Corning 365 silicon Emulsions).The silicon coating vial of the method manufacturing beyond any usefulness is above-mentioned and commercially available silicon coating vial all can use.
DTwPH combined vaccine according to the present invention is by being adsorbed onto diphtheria toxoid, tetanus toxoid and HBsAg stock solution on the alumina gel respectively, and above-mentioned solution mixed with the pertussis stock solution, the concentration of adjusting every kind of single antigen vaccine prepares to proper level.Diphtheria toxoid stock solution and tetanus toxoid stock solution are adsorbed in Fosfalugel (Yamanouchi), and the HBsAg stock solution is adsorbed in gel aluminum hydroxide.
According to single antigen vaccine concentration in the DTwPH combined vaccine of the present invention be: diphtheria toxoid 20-50Lf/mL, tetanus toxoid 4-20Lf/mL, pertussis 10-24OU/mL, HBsAg10-30 μ g/mL.The concentration of gel aluminum hydroxide is 15-35 μ gAl/mL.Total alumina gel concentration is 550-1000 μ gAl/mL.Term " concentration of gel aluminum hydroxide " or " concentration of alumina gel " refer to the concentration of aluminum in the alumina gel.
Fig. 1 is for amplifying the photo of 100X, the bordetella pertussis aggregation extent of gel aluminum hydroxide under the demonstration different hydro alumina concentration.When gel aluminum hydroxide and bordetella pertussis mix, because of electrostatic attraction is assembled.In order to make the gathering between gel aluminum hydroxide and the bordetella pertussis reduce to minimum, determine that the concentration of gel aluminum hydroxide is considered to a key factor.When the bordetella pertussis final concentration is 24OU/mL, when gel aluminum hydroxide concentration 35 μ gAl/mL, do not observe gathering, shown in Fig. 1 (a).When gel aluminum hydroxide concentration was not less than 35 μ gAl/mL, gel aluminum hydroxide and bordetella pertussis began to assemble, and aggregation extent increases the weight of when aluminium hydroxide concentration is not less than 200 μ gAl/mL, shown in Fig. 1 (b).
Therefore, the gel aluminum hydroxide concentration preferable range of absorption HBsAg is at 15-35 μ gAl/mL (based on final combined vaccine composition).The diphtheria toxoid adsorbent solution is not charged or electronegative under neutral pH, positively charged under faintly acid pH.Simultaneously, the bordetella pertussis adsorbent solution is electronegative at the neutral pH lower surface.Therefore, when pertussis stock solution and diphtheria toxoid adsorbent solution (are measured Zeta-Meter system 3.0, NJB PACIFIC INC., the U.S.) with pH meter when pH adjusts to scope 6.5-7.5, can prevent gathering in the mixed solution by electrostatic repulsion.
When the pH with diphtheria toxoid, tetanus toxoid and HBsAg adsorbent solution and bordetella pertussis solution is adjusted into the 6.5-7.5 scope and they is mixed, final DTwPH combined vaccine composition pH is 6.5-7.5, therefore can prevent the gathering between each antigen vaccine.
Meet preparation standard biology (korean foods and drug control, 1999) of each single antigen vaccine by the DTwPH combined vaccine according to the present invention of method for preparing, have the toxicity of reduction.In addition, the immunogenicity of HBsAg is stable in the DTwPH combined vaccine.Here, immunogenic stability refers in the immunogenicity of 4 ℃ of storage HBsAg after 3 months and initial level remains unchanged or slightly increase, does not significantly reduce.
The gathering that vaccine is not expected in storage or to the adhesion of vial wall, can be by adding high-viscosity material (as the gelatin of 0.1-0.7w/v%) to common combined vaccine or in according to DTwPH combined vaccine of the present invention, or preserve by the vial that uses silicon coating and to prevent.Like this, common combined vaccine or DTwPH combined vaccine according to the present invention remain white stable suspension in storage.Here, it is 2-8 ℃ cold room that storage-stable refers to be housed in temperature, and combined vaccine remains white suspension, and does not have the gathering of vaccine and to the adhesion of storage bottle.
The chart summary
Fig. 1 is for amplifying the photo of 100X, the bordetella pertussis aggregation extent of gel aluminum hydroxide under the demonstration different hydro alumina concentration.
Fig. 2 is the protein hybridization photo, shows the degree of absorption of diphtheria toxoid adsorbent solution.
Fig. 3 is the photo of the amplification 100X of optical microscope shooting, shows the influence of the vial storage of use silicon coating to aggregation extent in the DTwPH combined vaccine.
Embodiment
The present invention will describe in more detail by the following example.The following example is used to set forth, rather than is used for limiting the scope of the invention.
Embodiment 1: preparation diphtheria toxoid stock solution
The diphtheria toxoid stock solution that the present invention uses is by cultivating diphtheria corynebacterium Park Willium strain preparation.Adding final concentration to the culture supernatant that produces is 6% formalin, and places at 36 ℃ and to be used for detoxification in 28 days, then by ultrafiltration, dialysis, ammonium sulfate precipitation, gel permeation chromatography purification, and dialysis acquisition diphtheria toxoid stock solution then.
Embodiment 2: preparation tetanus toxoid stock solution
The tetanus toxoid stock solution is by cultivating clostridium tetani Harvard strain preparation.Add final concentration and be 0.45% formalin in the culture fluid that produces, and place at 36 ℃ and to be used for detoxification in 28 days, then by microfiltration, ultrafiltration, dialysis, ammonium sulfate precipitation, gel permeation chromatography purification, dialysis obtains the tetanus toxoid stock solution then.
Embodiment 3: preparation whole cell pertussis stock solution
The pertussis stock solution prepares by cultivating bordetella pertussis.With the medium centrifugal that produces.The pertussis precipitation that obtains is suspended in the saline and at 56 ℃ handles deactivation in 30 minutes, to obtain suitable pertussis stock solution.
Embodiment 4: preparation HBsAg stock solution
HBsAg prepares by cultivating reorganization multiple-shaped nuohan inferior yeast (Hansenula polymorpha).Carry out cell extraction after the culture homogenate that produces, then through ion chromatography, ultrafiltration, dialysis, super from obtaining suitable HBsAg stock solution with gel permeation chromatography.
Embodiment 5: preparation diphtheria toxoid and tetanus toxoid adsorbent solution
Diphtheria toxoid and tetanus toxoid stock solution are adsorbed in Fosfalugel (Yamanouchi) (pH5.5) by mixing, and dilute with isotonic saline solution when needing.Can suitably adjust final albumen of adsorbent solution and toxoid concentration.In this embodiment, the concentration of diphtheria toxoid solution is adjusted to about 280Lf/mL, and the concentration of tetanus toxoid solution is adjusted to about 50Lf/mL.Corresponding adsorbent solution pH is 5.5-6.0.Can be observed diphtheria toxoid by high performance liquid chromatography (HPLC) and protein hybridization analysis is adsorbed on the Fosfalugel (Yamanouchi).The protein hybridization analysis the results are shown in Fig. 2.In Fig. 2, swimming lane 1 is a molecular weight marker, and swimming lane 2,3 and 4 is respectively the diphtheria toxoid solution of 0.025Lf/mL, 0.05Lf/mL and 0.10Lf/mL.Swimming lane 5 is the centrifugal supernatant of diphtheria toxoid adsorbent solution (diphtheria toxoid that is equivalent to 100% desorbing) of 0.4Lf/mL.It is in 2-8 ℃ the cold room that the diphtheria toxoid of preparation and tetanus toxoid adsorbent solution are housed in temperature.
Embodiment 6: preparation HBsAg adsorbent solution
In order to increase immunogenicity, HBsAg is adsorbed in gel aluminum hydroxide.Mix HBsAg stock solution and gel aluminum hydroxide, the HBsAg stock solution is adsorbed on the gel, dilute if desired and with isotonic saline solution.The concentration of gel aluminum hydroxide is adjusted into 60 μ gAl/mL, and the concentration of HBsAg stock solution is adjusted into 72 μ g/mL.Can be observed HBsAg by HPLC and protein hybridization analysis is adsorbed on the gel aluminum hydroxide.It is in 2-8 ℃ the cold room that the HBsAg adsorbent solution of preparation is housed in temperature.
Embodiment 7: preparation DTwPH combined vaccine
The adsorbent solution and the pertussis stock solution (192OU/mL) of preparation in embodiment 5 and 6 are mixed, obtain final DTwPH combined vaccine composition.For the gathering that prevents to take place owing to electrostatic attraction in pertussis stock solution and the adsorbent solution mixture, the pH with each solution before mixing transfers to 7.1.
At first, diphtheria toxoid adsorbent solution and tetanus toxoid adsorbent solution mix, and HBsAg adsorbent solution and deactivation pertussis stock solution add preparation DTwPH combined vaccine composition in this mixture.Next Fosfalugel (Yamanouchi) is added in the DTwPH combined vaccine composition, adjust total alumina gel concentration to the expection level.The concentration of each stock solution can be by adding saline or phosphate buffer adjustment.The thimerosal that adds 0.05-0.1%mg/mL is as antiseptic.The 2-phenyl phenol that then adds 3-6mg/mL if desired.It is 7.1 composition that final DTwPH combined vaccine composition has the pH shown in the table 1.
Table 1
| Composition | Quantity |
| Diphtheria toxoid (Lf/mL) | 40 |
| Tetanus toxoid (Lf/mL) | 15 |
| Bordetella pertussis (OU/mL) | 24 |
| HBsAg(μg/mL) | 24 |
| Gel aluminum hydroxide (μ gAl/mL) | 20 |
| Fosfalugel (Yamanouchi) (μ gAl/mL) | 780 |
The immunogenicity of HBsAg test in the EXPERIMENTAL EXAMPLE 1:DTwPH combined vaccine
For the immunogenicity of HBsAg in the DTwPH combined vaccine of measuring among the embodiment 7 preparation, this combined vaccine of the subcutaneous immunity of ICR mouse web portion in 30 4-5 ages in week.The geometric average antibody titre (Ab GMT) of this immunogenicity test and the mice percent with potent antibodies titre are measured in the back blood drawing of 4 weeks.At this, Ab GMT refers to be not less than the geometric mean of the mouse antibodies titre of 1mIU/mL, and the mice percent with potent antibodies titre refers to that mice that antibody titer is not less than 10mIU/mL accounts for the percent in all experiment mices.
As simultaneously treated matched group, mice is formed three DTwP combined vaccines (40Lf/mL diphtheria toxoid, 15Lf/mL tetanus toxoid and 24OU/mL bordetella pertussis) and the 0.5-mL single antigen Hepatitis B virus vaccine (24 μ g/mL) identical with embodiment 1 at the subcutaneous immune 0.5-mL antigen of different loci, with above-described same procedure HBsAg is tested capable immunogenicity then.
As shown in table 2, the vaccine independent with immunity simultaneously is the same, and the DTwPH combined vaccine has shown stable HBsAg immunogenicity (at Ab GMT and the mice percent with potent antibodies titre).
Table 2
| The DTwPH combined vaccine | Immune simultaneously independent vaccine | |
| The immunogenicity Ab GMT (mIU/mL) of HBsAg | 372 | 354 |
| Mice percent (%) with potent antibodies titre | 90 | 93 |
The potency test of EXPERIMENTAL EXAMPLE 2:DTwPH combined vaccine
Among the embodiment 7 in the DTwPH combined vaccine of preparation diphtheria toxoid, tetanus toxoid, whole cell pertussis and HBsAg potency test separately render a service according to the bio-pharmaceutical of korean foods and drug control and require (1999,122-130 diphtheria, tetanus, pertussis absorption combined vaccine; With the 310-313 Hepatitis B virus vaccine) carry out.
The result is diphtheria toxoid 2-4 unit/mL, tetanus toxoid 4 units/mL or higher, whole cell pertussis 10IU/mL.The effectiveness of HBsAg is equal to or greater than reference material (using the standard of green cross company).Obviously, the DTwPH combined vaccine meets the effectiveness requirement of bio-pharmaceutical.
Embodiment 8: the use of gelatin
In order to increase the viscosity of the DTwPH combined vaccine composition of preparation among the embodiment 7, adding final concentration is the gelatin of 0.6w/v%.The result does not observe the gathering of vaccine after 3 months and to the adhesion of vial wall 2～8 ℃ of storages.
Embodiment 9: the use of silicon coating vial
The 0.5ml DTwPH combined vaccine composition branch of preparation among the embodiment 7 is installed in the vial (3mL) and vial that does not have coating of a silicon coating, or for more time 2-8 ℃ of storage 3 months.The result does not observe the adhesion of vaccine to the vial wall of silicon coating.After acutely shaking these two bottles, observe the steady statue of vaccine combination at 100X (Axioplan, Carl Zeiss Co., Germany) with microscope.The DTwPH combined vaccine composition that is housed in the silicon coating vial has been kept initial homogenizing dispersity in storage, shown in Fig. 3 (a).On the contrary, in not having the vial of silicon coating, observed the gathering of DTwPH combined vaccine, shown in Fig. 3 (b).
Industrial usability
As mentioned above, the gathering of vaccine and can be by preparation DTwPH combined vaccine behind the pH that adjusts gel aluminum hydroxide concentration and each adsorbent solution and pertussis stock solution to the adhesion of storage vial, or by in the DTwPH combined vaccine, adding gelatin, or use the vial of silicon coating to preserve to prevent.
Diphtheria toxoid and tetanus toxoid are adsorbed onto on the Fosfalugel (Yamanouchi), HBsAg is adsorbed onto on the gel aluminum hydroxide, and prepare the DTwPH combined vaccine after adjusting each constituent concentration, the immunogenicity that can keep HBsAg in the DTwPH combined vaccine, the effectiveness of each composition can satisfy the requirement (korean foods and medication management, 1999) of preparation standard biology.
In addition, other single antigen vaccine such as haemophilus influenzae type b (Hib) vaccine or PKV can add in the DTwPH combined vaccine goods, or mix with it immediately before the inoculation at the same time.
Claims (11)
1. a DTwPH (diph/tet-pertussis-hepatitis) combined vaccine, wherein diphtheria toxoid and tetanus toxoid are adsorbed in Fosfalugel (Yamanouchi), and hepatitis B surface antigen is adsorbed in gel aluminum hydroxide.
2. according to the DTwPH combined vaccine of claim 1, final pH is 6.5-7.5.
3. according to the DTwPH combined vaccine of claim 1 or 2, wherein the concentration range of gel aluminum hydroxide is 15-35 μ gAl/mL.
4. according to the DTwPH combined vaccine of claim 1, wherein the concentration range of diphtheria toxoid is that the concentration range of 20-50Lf/mL, tetanus toxoid is that 4-20Lf/mL, pertussal concentration range are that the concentration range of 10-24OU/mL, hepatitis B surface antigen is 10-30 μ g/mL, and total alumina gel concentration range is 550-1000 μ gAl/mL.
5. the combined vaccine of claim 1, it contains the gelatin of 0.1-0.7w/v% consumption.
6. according to the combined vaccine of claim 5, wherein the amount ranges of gelatin is 0.5-0.7w/v%.
7. prepare the method for DTwPH (diph/tet-pertussis-hepatitis) combined vaccine, this method comprises:
The Fosfalugel (Yamanouchi) solution of absorbed diphtheria toxoid is mixed (a) with the Fosfalugel (Yamanouchi) solution of adsorbed tetanus toxoid; With
The mixture that will obtain in step (a) mixes (b) with the gel aluminum hydroxide solution and the deactivation pertussis stock solution of absorption hepatitis B surface antigen.
8. according to the method for claim 7, wherein DTwPH combined vaccine final pH is 6.5-7.5.
9. according to the method for claim 7 or 8, wherein the concentration range of gel aluminum hydroxide is adjusted to 15-35 μ g/mL in the DTwPH combined vaccine.
10. method according to Claim 8, wherein the concentration range of the diphtheria toxoid concentration range that is adjusted to 20-50Lf/mL, tetanus toxoid is adjusted to the concentration range that 4-20Lf/mL, pertussal concentration range be adjusted to 10-24OU/mL, hepatitis B surface antigen and is adjusted to 10-30 μ g/mL, and total alumina gel concentration range is adjusted to 550-1000 μ gAl/mL.
11. the combined vaccine storage practice of a claim 1, wherein the combined vaccine packing of claim 1 and preserving in the vial of silicon coating.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2000-0038164A KR100385711B1 (en) | 2000-07-05 | 2000-07-05 | The quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis b surface antigen and the preparation thereof |
| KR2000/38164 | 2000-07-05 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1383384A CN1383384A (en) | 2002-12-04 |
| CN1211121C true CN1211121C (en) | 2005-07-20 |
Family
ID=19676257
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB018019250A Expired - Fee Related CN1211121C (en) | 2000-07-05 | 2001-07-05 | Quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis B surface antigen, and prepn. method thereof |
Country Status (4)
| Country | Link |
|---|---|
| KR (1) | KR100385711B1 (en) |
| CN (1) | CN1211121C (en) |
| AU (1) | AU2001269569A1 (en) |
| WO (1) | WO2002005846A1 (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0835663B1 (en) | 1992-05-23 | 2009-09-30 | GlaxoSmithKline Biologicals S.A. | Combined vaccines comprising Hepatitis B surface antigen and other antigens |
| KR100385711B1 (en) * | 2000-07-05 | 2003-05-27 | 녹십자백신 주식회사 | The quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis b surface antigen and the preparation thereof |
| ES2649048T3 (en) | 2002-11-01 | 2018-01-09 | Glaxosmithkline Biologicals S.A. | Drying procedure |
| GB0505518D0 (en) | 2005-03-17 | 2005-04-27 | Chiron Srl | Combination vaccines with whole cell pertussis antigen |
| TW200806315A (en) * | 2006-04-26 | 2008-02-01 | Wyeth Corp | Novel formulations which stabilize and inhibit precipitation of immunogenic compositions |
| PE20100365A1 (en) | 2008-10-24 | 2010-05-21 | Panacea Biotec Ltd | NEW COMBINATION VACCINES WITH WHOLE CELL COUGH AND METHOD FOR THEIR PREPARATION |
| PL3170508T3 (en) | 2010-06-04 | 2020-04-30 | Wyeth Llc | Vaccine formulations |
| FR2966044B1 (en) * | 2010-10-18 | 2012-11-02 | Sanofi Pasteur | METHOD FOR CONDITIONING A VACCINE CONTAINING AN ALUMINUM ADJUVANT |
| CN104487086B (en) * | 2012-07-07 | 2019-08-30 | 巴拉特生物技术国际有限公司 | Non-animal derived nonalcoholic vaccine composition and preparation method thereof |
| CN104341499B (en) * | 2014-11-06 | 2017-05-24 | 遵义医学院 | Preparation process of tetravalent yolk antibody preparation for resisting measles and bronchocephalitis, diphtheritis and lockjaw |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2322057A1 (en) * | 1973-05-02 | 1974-11-21 | Porsche Ag | DEVICE FOR CATALYTIC AFTER-BURNING OF EXHAUST GASES OF A MULTICYLINDRICAL COMBUSTION MACHINE |
| JP2743177B2 (en) * | 1987-04-24 | 1998-04-22 | 財団法人阪大微生物病研究会 | Method of culturing pertussis, pertussis toxoid and combination vaccine thereof |
| KR920009729A (en) * | 1990-11-26 | 1992-06-25 | 전금자 | Pottery manufacturing method that generates far infrared rays |
| AU2088992A (en) * | 1992-05-05 | 1993-11-11 | Research Foundation For Microbial Diseases Of Osaka University, The | Stabilized live vaccine |
| EP0835663B1 (en) * | 1992-05-23 | 2009-09-30 | GlaxoSmithKline Biologicals S.A. | Combined vaccines comprising Hepatitis B surface antigen and other antigens |
| JPH10131967A (en) * | 1996-10-25 | 1998-05-22 | Seiko Epson Corp | Bearing and motor |
| GB9806456D0 (en) * | 1998-03-25 | 1998-05-27 | Smithkline Beecham Biolog | Vaccine composition |
| RU2130778C1 (en) * | 1998-07-31 | 1999-05-27 | Акционерное общество закрытого типа НПК "Комбиотех Лтд" | Mixed vaccine for immune prophylaxis of viral hepatitis b, tetanus, diphtheria and whooping cough |
| KR100385711B1 (en) * | 2000-07-05 | 2003-05-27 | 녹십자백신 주식회사 | The quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis b surface antigen and the preparation thereof |
-
2000
- 2000-07-05 KR KR10-2000-0038164A patent/KR100385711B1/en active IP Right Grant
-
2001
- 2001-07-05 AU AU2001269569A patent/AU2001269569A1/en not_active Abandoned
- 2001-07-05 WO PCT/KR2001/001153 patent/WO2002005846A1/en active Application Filing
- 2001-07-05 CN CNB018019250A patent/CN1211121C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| KR100385711B1 (en) | 2003-05-27 |
| CN1383384A (en) | 2002-12-04 |
| KR20020005081A (en) | 2002-01-17 |
| WO2002005846A1 (en) | 2002-01-24 |
| AU2001269569A1 (en) | 2002-01-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1211121C (en) | Quadrivalent combination vaccine including diphtheria toxoid, tetanus toxoid, whole cell pertussis and hepatitis B surface antigen, and prepn. method thereof | |
| US4455297A (en) | Method for producing pertussis toxoid | |
| CN104689309A (en) | Separated and purified acellular pertussis-diphtheria-tetanus, b-type haemophilus influenzae and A-group and C-group meningococcus combined vaccine and preparation method thereof | |
| EP2075005B1 (en) | Ipv-dpt vaccine | |
| US20080193478A1 (en) | Inactivated Poliomyelitis Vaccine Derived From Sabin Strain Of Polio Virus | |
| CN1712068A (en) | Diploid-cell rabies vaccine and purified rabies vaccine, freeze-drying preparation and water injection thereof | |
| CN1228086C (en) | Preparation process for combined vaccine | |
| WO2024125100A1 (en) | Bivalent inactivated ev71-ca16 vaccine, method for preparing same, and use thereof | |
| CN1686540A (en) | Preparation of tetravalent wheel shaped virus inactivated vaccine and application | |
| CN100467062C (en) | Method of production of purified hepatitis A virus particles and vaccine preparation | |
| CN1617923A (en) | Method of large scale production of hepatitis a virus | |
| WO2000057904B1 (en) | Attenuated dengue-3 virus vaccine | |
| CN1218750C (en) | Recombinant hepatitis B BCG vaccine combined vaccine and its preparation method | |
| CN1843507A (en) | Mumps virus ingredient vaccine for human, and its preparation method and uses | |
| CN1308547A (en) | Novel compositions | |
| JP2019527191A (en) | Immune enhancer, foot-and-mouth disease inactivated vaccine, and method for producing the same | |
| CN1299768C (en) | Diploid cell cerebritis B vaccine and purified cerebritis B vaccine, dosage form freeze-drying and water injection | |
| CN1228085C (en) | Forest cerebritis purifying vaccinum | |
| CN1216985C (en) | Hepatitis A virus strain, method for preparing hepatitis A inactivated vaccine and obtained vaccine | |
| CN1990041A (en) | Kidney syndrome hemorrhagic fever Vero cell bivalent purified vaccine and industrialized producing process thereof | |
| EP3768821A1 (en) | Methods of cultivating bordetella species | |
| CN1314449C (en) | Combined vaccine composed of viral of Japanese b encephalitis and vaccine of brain fever cocci | |
| CN1305527C (en) | Vaccine for treating hepatitis B, and its prepn. method | |
| CN115475240A (en) | Application of artemisia rupestris L crude polysaccharide cultured in Xinjiang as foot-and-mouth disease inactivated vaccine adjuvant | |
| CN1401389A (en) | Hepatitis B vaccine preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| 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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20050720 Termination date: 20200705 |