CN106075428A - A kind of immunogenic composition and preparation method thereof - Google Patents
A kind of immunogenic composition and preparation method thereof Download PDFInfo
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Abstract
The invention provides a kind of immunogenic composition, belong to technical field of biological product preparation, this immunogenic composition contains polio antigen and the b type hemophilus influenza GL-PP conjugate of inactivation.It also can comprise hepatitis B antigen and physiologically acceptable carrier.Present invention also offers the preparation method of described immunogenic composition.The immunogenic composition of the present invention can prevent not exist between infecting of multiple pathogens, and these antigens the phenomenon interfered simultaneously, and the immunogenicity that corresponding immunogenicity excites compared with antigen alone will not reduce；And use the immunogenic composition of the present invention can substantially simplify vaccination program, improve inoculation efficiency and reduce cost.
Description
Technical field
The present invention relates to technical field of biological product preparation, in particular it relates to a kind of immunogenic composition and preparation thereof
Method.
Background technology
Poliomyelitis is to be infected, by virus, the highly infective disease that people causes.Poliovirus can infect human body
Nervous system, and patient limb can be caused within a few hours to benumb, cause life-long disabilities.Poliomyelitis can be at all age brackets
Occurring in crowd, but be apt to occur in the infant less than 5 years old, therefore this disease is also called " poliomyelitis ".Poliovirus
Belong to the enterovirus genus of Picornaviridae, I, II and type III can be divided into according to serum group system, all can so that
Sick.Up to the present medical circle does not the most treat poliomyelitic specific drug, but poliomyelitis can be by injection epidemic disease
Seedling reaches good preventive effect.
The poliomyelitis vaccine (IPV) of inactivation is succeeded in developing in the fifties in last century by doctor Salk at first.1954
Having carried out clinical trial in the U.S., next year clinical test results proves that vaccine has good safety and protectiveness,
Large-scale inoculation has been carried out subsequently movable in the U.S..The poliomyelitis vaccine (OPV) that another attenuation is administered orally is won by Sabin
Scholar succeeded in developing in the sixties in last century, and worldwide used rapidly, contribute to heavily fortified point for preventing and treating Incidence of Poliomyelitis
Strength amount, but, owing to OPV is attenuated vaccine, during taking, there is certain potential safety hazard, therefore, OPV is progressively gone out
The poliomyelitis vaccine IPV lived replaces.World Health Organization (WHO) (WHO) starts whole world poliomyelitis and eliminates action, encourages
Developing country uses the IPV of multivalence or unit price in planned immunization, thus thoroughly blocks poliovirus in crowd
Infection.The poliomyelitis vaccine of inactivation can be divided into two classes: the inactivated vaccine prepared by street strain and being prepared by attenuated strain
Inactivated vaccine (sabin-IPV).The popularization that significantly limit in developing country of street strain is used to make owing to the former produces IPV
With, especially epoch after Poliomyelitis Eradication virus, WHO can the tightened up use limiting street strain, therefore sabin-IPV
Vaccine just becomes a kind of new product replacing OPV vaccine to promote the use of in developing country.
Hib (b type hemophilus influenza) is the symbiotic bacteria that child's nasopharynx part is common, can cause children Streptococcus and meninges
Inflammation, in the area not yet carrying out extensive Hib vaccination, Hib disease is a main public health problem.The most at least
Having 3,000,000 several cases to occur, about 38.6 die ten thousand deaths dies.Hib morbidity is more common in developing country with death, and Disease Spectrum is at 4-18
Monthly age child is the most serious, but is less than for 3 monthly ages and the most occasionally has morbidity more than 5 years old child.Not immunity crowd, within Hib is 1 years old
The Etiological of the non-popular bacterial meningitis of child.Even if giving enough antibiotic therapies in time, the Hib brain of 3-20%
Film inflammation patient still can be dead.
Hib is infected maximally effective preventive measure is intramuscular injection Hib coupling vaccine, can be substantially reduced pneumonia and meningitis
Incidence probability.Although Hib capsular polysaccharide has certain immunogenicity, but owing to child immune system is grown unsound, to many
The immunne response of sugar is more weak, and therefore, immunity after need to carrying out puting together by Hib capsular polysaccharide and carrier protein, in exciting infants
TD immunization route, to strengthen the infant immunne response to polysaccharide.
Poliomyelitis vaccine and Hib combined vaccine, with as the vaccine for infant planned immunization, have and are subject to widely
The crowd of kind, and the age of inoculation group is more similar.These two kinds of vaccines are multi-agent immunity, are needed repeatedly to inoculate by kind of a crowd, time-consuming
Arduously.It would therefore be highly desirable to need a kind of combined vaccine, save by kind of a number of times while not reducing the two immune effect, during saving
Between and human resources.
Summary of the invention
In order to solve problems of the prior art, it is an object of the invention to provide a kind of containing inactivation poliomyelitis
Virus and the immunogenic composition of Hib GL-PP conjugate.
In order to realize the object of the invention, the invention provides a kind of immunogenic composition, described immunogenic composition
Polio virus antigens and Hib GL-PP conjugate including inactivation.
The poliovirus of described inactivation is selected from sabin strain, comprises sabin I type, sabin II type and sabin III
Type.
Described immunogenic composition contains sabin I type antigen 2.5-20D unit/person-portion, sabin II type antigen 5-
40D unit/person-portion, sabin type III antigen 5-70D unit/person-portion, Hib GL-PP conjugate 1-15 μ g/ person-portion.
Preferably, described immunogenic composition contains sabin I type antigen 5-15D unit/person-portion, and sabin II type resists
Former 10-30D unit/person-portion, sabin type III antigen 1 0-40D unit/person-portion, Hib GL-PP conjugate 5-10 μ g/ people
Part.
Further, described immunogenic composition can also be containing hepatitis B antigen after purification.Preferably, containing purification
After hepatitis B antigen be 5-40 μ g/ person-portion.
It is highly preferred that described immunogenic composition comprises hepatitis B antigen 15-30 μ g/ person-portion.
Further, foregoing immune originality compositions can also comprise physiologically acceptable carrier, and comprising can be with b type influenza
The physiologically acceptable carrier protein that haemophilus polysaccharide antigen is puted together, such as the tetanus toxoid carrier of 10-50 μ g/ person-portion,
Also selected from diphtheria toxoid, diphtheria CRM197 albumen and meningitis outer membrane protein or other carrier any of.
Bacterial polysaccharides uses known technology with the coupling method of carrier protein, including using 1-cyano group-4-dimethylamino pyrrole
After pyridine tetrafluoroborate (CDAP) activated polysaccharide antigen covalently bound with carrier protein, also include use reductive amination method process many
Sugar and the coupling of carrier protein, or use after cyanogen bromide-activated polysaccharide the method with carrier protein couplet.
Further, described immunogenic composition can also comprise adjuvant.
Preferably, described adjuvant is aluminium salt.
It is highly preferred that described aluminium salt is aluminum phosphate or aluminium hydroxide, most preferably aluminium hydroxide.
Further, the dosage form of foregoing immune originality compositions be injection, lyophilized preparation, injection, capsule, tablet or
Pill.
Present invention also offers described immunogenic composition and in preparation prevention or treat poliomyelitis or meningitis medicine
Application in thing.
Present invention also offers the application in preparing vaccine of the above-mentioned immunogenic composition.
The preparation method of foregoing immune originality compositions median ridge ash antigen is by Technology manufacture ripe in industry
, including cultivation, results, purification and the inactivation of three kinds of serotype Polio virus, the cellular matrix that wherein Polio virus is cultivated can
To select human diploid cell such as 2BS cell or passage cell such as Vero cell, training method can select cell factory or
Person's bioreactor；The purification of ridge ash antigen can include multistage clarification filtration, concentration, sieve chromatography and ion-exchange chromatography；
The inactivation of ridge ash antigen uses formaldehyde, and method meets WHO to requirement to inactivation step in inactivation OPV guideline.
In foregoing immune originality compositions, the preparation method of Hib GL-PP conjugate is by technique ripe in industry
Technology manufactures, and by obtaining the cultivation of Hib bacterium and purification, including the cultivation in fermentation tank of the Hib bacterium, thalline is carried out good fortune
The process of that Malin's process, centrifugal concentrating and a series of chemical method obtains capsular polysaccharide after purification, then by a kind of life
Reason acceptable carriers albumen, coupling method based on a kind of above-mentioned polysaccharide and carrier protein obtains Hib capsular polysaccharide conjugate.
Preparation method meets 2010 editions related requests preparing Hib antigen conjugates of Chinese Pharmacopoeia.The Hib polysaccharide egg that the present invention uses
In white conjugate, Hib polysaccharide antigen is 1:2-2:1 with the mass ratio of carrier protein.
In foregoing immune originality compositions, the preparation method of hepatitis B antigen is by Technology manufacture ripe in industry
, including expression and the purification of recombination hepatitis B antigen, wherein recombination hepatitis B expression vector can be Hansenula yeast or medicated beer ferment
Mother, is cultivated by bacterial fermentation tank and expresses hepatitis B antigen；The results of hepatitis B antigen and purification can include grinding brokenly bacterium, PEG sinks
Shallow lake, ultrafiltration concentration, sieve chromatography, ion-exchange chromatography and centrifugal desalination.Preparation method meets Chinese Pharmacopoeia 2010 editions to second
Related request prepared by liver antigen.
The preparation of immunogenic composition: the Polio virus antigens stock solution of inactivation is puted together with Hib GL-PP
Thing, the hepatitis B antigen of purification are stirred mixing, and control the pH value of mixed liquor between 6-7.If antigen mixture comprises
Adjuvant, then carry out mix and blend after antigen and adjuvant being adsorbed, and control the pH value of mixed liquor between 6-7；Described go out
The Polio virus antigens stock solution lived contains sabin I type antigen 2.5-20D unit/person-portion, sabin II type antigen 5-
40D unit/person-portion, sabin type III antigen 5-70D unit/person-portion, b type hemophilus influenza GL-PP conjugate 1-15 μ
G/ person-portion；Hepatitis B antigen 5-40 μ g/ person-portion.
Preferably, the Polio virus antigens stock solution of described inactivation contains sabin I type antigen 5-15D unit/people
Part, sabin II type antigen 1 0-30D unit/person-portion, sabin type III antigen 1 0-40D unit/person-portion, the bloodthirsty bar of b type influenza
Granulose protein conjugate 5-10 μ g/ person-portion；Hepatitis B antigen 15-30 μ g/ person-portion.
The immunogenicity of aforementioned antigens:
The immunogenicity of immunogenic composition median ridge ash antigen is evaluated by the experiment of rat titer, simultaneously with inactivation
Ridge ash univalent vaccine contrasts.SD rat or Wistar rat is selected to carry out intramuscular injection according to 10 specifications often organized and exempt from
Epidemic disease, antigen mixture carries out 3 times of doubling dilutions according to people with dosage, is made into 5 groups of antigen mixtures altogether.After rat immunity the 3rd week
Strengthen once, within after immunity the 2nd week, take a blood sample, with virus residual titration experiment detection NAT.This type of neutralize experiment according to
2010 editions requirements of Chinese Pharmacopoeia are carried out.
In immunogenic composition, the immunogenicity of b type hemophilus influenza antigen is little by BALB/c mouse or NIH
The experiment of Mus titer is evaluated, and contrasts with b type hemophilus influenza combined vaccine simultaneously.Mice is according to 20 rule often organized
Lattice carry out injecting immune.Antigen mixture carries out immunity according to the injection volume of the antigen mixture of each 2.5 μ g polysaccharide.Little mouse's head
First carry out 0.85% sodium chloride solution injection, injected twice in the 1st, 14 days, took a blood sample through orbital vein with the 21st～28 day, use
ELISA method measures the IgG antibody of Hib.Titer experiment is carried out according to 2010 editions requirements of Chinese Pharmacopoeia.
In immunogenic composition, the immunogenicity of hepatitis B antigen is evaluated, simultaneously by the experiment of BALB/c mouse titer
Contrast with unit price reconstituted hepatitis B vaccine.Mice carries out injecting immune according to 20 specifications often organized, and takes a blood sample, adopt after 4-6 week
Measure hbv antibody by ELISA method, calculate median effective dose ED50.ED50 experiment according to Chinese Pharmacopoeia 2010 editions require into
OK.
The immunogenic composition of the present invention is by Polio virus antigens and b type hemophilus influenza polysaccharide antigen
Combine, be subject to kind of a crowd, while not reducing the two immune effect with the form immunity of Combined vaccine or more combined vaccine
Save by kind of a number of times, save time and human resources.And the production strain of Polio virus antigens of the present invention is for going out
The attenuated strain lived, compares with OPV it can be avoided that the risk of strain virulence reversion, is respectively provided with good safety to by kind person and environment
Property.Combined vaccine prepared by the immunogenic composition of the present invention comprises Polio virus antigens and HIB polysaccharide antigen, energy
Enough poliomyelitis well preventing to be caused by poliovirus and HIB and meningitic generation.And present invention research
Show, above-mentioned each antigen in immunity by not disturbing antigenicity and immune effect mutually after kind person, have good immunogenicity with
Safety.
Detailed description of the invention
Following example are used for illustrating the present invention, but are not limited to the scope of the present invention.Without departing substantially from present invention spirit
In the case of essence, the amendment that the inventive method, step or condition are made or replacement, belong to the scope of the present invention.As
The lower embodiment that we are adopted employs the technology of standard, unless otherwise detailed description, by those skilled in the art crowd week
Know and for routine techniques.Embodiment is illustrative, but is not limiting as the present invention.
The cultivation of embodiment 1Sabin poliovirus
The ridge ash strain sabin I type that thered is provided by U.S. ATCC, sabin II type, sabin type III, thin at 10 layers respectively
Adaptable growth continuous passage in born of the same parents factory Vero cell, use MEM culture medium to add 8% calf serum (v/v), put 37 DEG C of cultivations
Change serum-free M199 culture medium after case is cultivated 4 days, connect sabin strain I type, II type and type III respectively according to MOI=0.01 sick
Poison, gathers in the crops virus liquid after cultivating 3 days at 33 DEG C, uses CCID after clarified filtration50Method detection virus titer value.Above three type ridge ashes
Virus harvest liquid can be at 7-8LgCCID50In the range of/ml.Virus harvest liquid is simultaneously by D antigen ELISA double antibodies sandwich detection method
Detection D antigen value.The D antigenic content of above three type Polio virus harvest liquids is in the range of 40-110DU/ml.
The purification of embodiment 2Sabin poliovirus and inactivation
By in embodiment 1 obtain Polio virus harvest liquid carry out three grades of clarification filtrations, select 0.75 μm, 0.45 μm and
The filter membrane bag of 0.22 μm；Feed liquid after clarification filtration carries out two-stage ultrafiltration concentration, and the first order selects 100KD polyether sulfone film
Bag concentrates 40 times, and 100KD polyether sulfone film bag reconcentration 10 times is selected in the second level；Then the feed liquid after concentrating passes through molecular sieve
Chromatographic column, pillar filler is selected Sephadex CL-6B, is balanced with 0.1M PBS (pH 7.0) buffer, then proceedes to use
0.1M PBS (pH 7.0) eluting, collecting the 1st peak is virus peak.It is added to the virus liquid of collection put down with 0.1M PBS (pH 7.0)
In the chromatographic column equipped with DEAE Sepharose Fast Flow weighed, with 0.1M PBS (pH 7.0) eluting, collect the 1st and wash
De-peak is virus peak.This collection virus liquid inactivates 12 days with 37 DEG C of 1:4000 formaldehyde after being filtered by 0.22 μm, after through degerming
Poliovirus strain sabin I type, sabin II type, sabin type III antigen stock is obtained after filtration.
The cultivation of embodiment 3B type hemophilus influenza and polysaccharide purification
According to description to b type hemophilus influenza cultural method in the Chinese patent of Patent No. 200810125160,
Including 2 precultures on solid medium and the preculture of No. 1 fluid medium, then cultivate in fermentation tank and receive
Obtain.According to description to b type hemophilus influenza polysaccharide purification method in this patent, carry out at formalin including by harvest liquid
Reason, centrifugal antigen supernatant of collecting also concentrate, and carry out the process of a series of chemical method subsequently, it is thus achieved that the b type influenza of purification addicted to
Blood bacillus polysaccharide, wherein Hib GL-PP antigen is 1:2-2:1 with the mass ratio of carrier protein.
The preparation of embodiment 4 hemophilus influenza GL-PP conjugate
According in publication 200680023334.4 to b type hemophilus influenza GL-PP and diphtheria toxoid mutation
The description of the covalent bonding approach of CRM197 albumen, carries out activation processing, and a series ofization including adding hydrogen bromide to polysaccharide
B type hemophilus influenza polysaccharide and CRM197 are carried out coupling and form conjugate by method, then conjugate is carried out chromatography purification,
Obtain hemophilus influenza combined vaccinogen liquid.
The preparation of embodiment 5 hepatitis B antigen stock solution
Use and cultivate the method that Hansenula yeast ATCC No.26012 prepares recombination hepatitis B antigen, at shaking flask and bacterial fermentation
In tank, three grades of levels carry out gathering in the crops brokenly bacterium after cultivating Hansenula yeasts, homogenizer can be used to grind broken, then carry out sulfur ammonium precipitation and
Being concentrated by ultrafiltration, concentrated solution carries out chromatography purification, ion exchange DEAE carries out column chromatography, it is thus achieved that hepatitis B stock solution.
Embodiment 6 ridge ash and the preparation of HIB immunogenic composition
The HIB stock solution that the ridge ash antigen stock prepared in embodiment 2 is prepared with embodiment 4 is pressed finite concentration stirring mixed
Close, it is thus achieved that immunogenic composition, determine that pH value is in the range of 6-7.Compositions antigenic content is shown in Table 1.
The grey preparation with HIB immunogenic composition of the embodiment 7 ridge containing adjuvant
By in embodiment 2 prepare ridge ash antigen stock and embodiment 4 described in HIB GL-PP conjugate respectively with
The aluminum hydroxide solution equal-volume of 0.5mg/ml uniformly mixes, and absorption 30 minute is stirred at room temperature, and makes the antigen containing aluminium adjuvant half
Finished product, then by the ridge ash antigen containing adjuvant and HIB antigen equal-volume mix and blend, it is thus achieved that the vaccine containing aluminium adjuvant, determines
PH value is in the range of 6-7.Compositions antigenic content is shown in Table 1.
Table 1 ridge ash and HIB compositions antigenic content
Embodiment 8 ridge ash, HIB, the preparation of immunogenic composition of hepatitis B antigen
By the HIB GL-PP conjugate prepared in the ridge ash antigen stock prepared in embodiment 2, embodiment 4 and enforcement
The hepatitis B antigen stock solution prepared in example 5 presses finite concentration stirring mixing, it is thus achieved that immunogenic composition, determines that pH value is 6-7's
In the range of.In combined vaccine composition, each antigenic content is shown in Table 2.
Embodiment 9 contains the preparation of the immunogenic composition of the ridge ash of adjuvant, HIB, hepatitis B antigen
By the HIB GL-PP conjugate described in the ridge ash antigen stock described in embodiment 2, embodiment 4 and enforcement
Hepatitis B antigen stock solution described in example 5 uniformly mixes with the aluminum hydroxide solution equal-volume of 0.5mg/ml respectively, and suction is stirred at room temperature
Attached 30 minutes, make the antigen semi-finished product containing aluminium adjuvant, then by the above-mentioned antigen equal-volume mix and blend containing adjuvant, it is thus achieved that contain
The combined vaccine composition of aluminium adjuvant, determines that pH value is in the range of 6-7.In combined vaccine composition, each antigenic content is shown in Table 2.
Antigenic content in table 2 ridge ash, HIB, hepatitis B compositions
Embodiment 10 ridge ash, the immunogenicity of HIB conjugate composition
By SD rat respectively immunity embodiment 6 is obtained containing ridge ash and the immunogenic composition of HIB conjugate and
The immunogenic composition containing ridge ash and HIB conjugate antigen and aluminum hydroxide adjuvant that embodiment 7 obtains assesses SD rat
Immunogenicity to compositions median ridge ash.Rat carries out injecting immune according to 10 specifications often organized.Immunogenic composition is pressed
Carry out 3 times of doubling dilutions according to people with dosage, be made into 5 groups altogether.Select inactivation OPV as a control group, after rat immunity the 3rd
Zhou Jiaqiang once, takes a blood sample for after immunity the 2nd week, with virus residual titration experiment detection NAT and seropositive conversion.Neutralize
Selecting Sabin I, II and type III by virus, various titre is 100CCID50.Positive control cell selects Vero cell.In
7 days, and observation of cell pathological changes is carried out at 36 DEG C with test.Result shows that each group of rat blood serum all has sabin I, II and III
The neutralization of type virus and seropositive conversion, illustrate the immunogenic composition inoculation rat energy that embodiment 6 and embodiment 7 prepare
Enough obtain good immunogenicity, show simultaneously and the immunogenicity of this immunogenic composition and matched group inactivate OPV
Immunogenicity suitable, experimental result is shown in Table 3.
By the grey and IMMUNOGENIC COMPOSITION of HIB conjugate antigen containing ridge that NIH mice immunity embodiment 6 respectively is prepared
The compositions containing ridge ash and HIB conjugate antigen and aluminum hydroxide adjuvant that thing and embodiment 7 prepare assesses NIH mice to group
The immunogenicity of HIB in compound.Selecting HIB combined vaccine as a control group, mice is injected according to 20 specifications often organized
Immunity (containing the group of 10 comparisons/often).Immunogenic composition carries out immunity according to each 2.5 μ g injection volumes.First mice is carried out
0.85% sodium chloride solution injection, injected twice in the 1st, 14 days, took a blood sample through orbital vein with the 21st～28 day, use ELISA method
Measure the IgG antibody of HIB.It is right that the immunogenic composition that result display embodiment 6 and embodiment 7 prepare meets on Chinese Pharmacopoeia
The relevant regulations of HIB titer, shows immunogenicity and the immunogenicity phase of matched group HIB combined vaccine of this anti-compositions simultaneously
When, experimental result is shown in Table 3.
Embodiment 11 ridge ash, HIB conjugate, the immunogenicity of hepatitis B compositions
Resisted containing ridge ash antigen, HIB GL-PP conjugate, hepatitis B by what SD rat immunity embodiment 8 respectively was prepared
Former compositions and embodiment 9 prepare containing ridge ash antigen and HIB GL-PP conjugate, hepatitis B antigen and aluminum hydroxide adjuvant
Compositions assess SD rat to compositions median ridge ash immunogenicity.Rat carries out injection according to 10 specifications often organized and exempts from
Epidemic disease.Immunogenic composition carries out 3 times of doubling dilutions according to people with dosage, is made into 5 groups altogether.Select inactivation OPV as right
According to group, within after rat immunity the 3rd week, strengthen once, within after immunity the 2nd week, take a blood sample, by virus residual titration experiment detection neutralizing antibody effect
Valency and seropositive conversion.Neutralizing and select Sabin I, II and type III by virus, various titre is 100CCID50.Positive control is thin
Born of the same parents select Vero cell.Neutralization test carries out 7 days, and observation of cell pathological changes at 36 DEG C.Result shows that each group of rat blood serum all has
The neutralization viral to sabin I, II and type III and seropositive conversion, illustrate the immunogen that embodiment 8 and embodiment 0 prepare
Then compositions has good immunogenicity on rat, shows in the immunogenicity of this antigen mixture and matched group simultaneously and goes out
The immunogenicity of OPV alive is suitable, and experimental result is shown in Table 3.
By NIH mice respectively immunity embodiment 8 prepared containing ridge ash, HIB polypeptide protein conjugate, hepatitis B antigen
Immunogenic composition and embodiment 9 prepare containing ridge ash, HIB conjugate, hepatitis B antigen and the combination of aluminum hydroxide adjuvant
Thing assesses NIH mice to the immunogenicity of HIB in compositions.Selecting HIB combined vaccine as a control group, mice is according to 20
The specification often organized carries out injecting immune (containing the group of 10 comparisons/often).Immunogenic composition is carried out according to each 2.5 μ g injection volumes
Immunity.First mice carries out 0.85% sodium chloride solution injection, injects twice in the 1st, 14 days, quiet through eye socket with the 21st～28 day
Arteries and veins is taken a blood sample, and measures the IgG antibody of HIB by ELISA method.The immune principle compositions that result display embodiment 8 and embodiment 9 prepare
Meet the relevant regulations to HIB titer on Chinese Pharmacopoeia, show immunogenicity and the matched group HIB knot of this antigen mixture simultaneously
Close the immunogenicity phase of vaccine, when experimental result is shown in Table 3.
Resisted containing ridge ash, HIB polypeptide protein conjugate, hepatitis B by what BALB/c mouse immunity embodiment 8 respectively was prepared
Former immunogenic composition and embodiment 9 prepare containing ridge ash, HIB conjugate, hepatitis B antigen and the group of aluminum hydroxide adjuvant
Compound assesses BALB/c mouse to the immunogenicity of hepatitis B virus in immunogenic composition.Select reconstituted hepatitis B vaccine conduct
Matched group, mice carries out injecting immune according to 20 specifications often organized, and takes a blood sample after 4-6 week, uses ELISA method to measure hepatitis B and resists
Body, calculates median effective dose ED50.It is right that the immunogenic composition that result display embodiment 8,9 prepares meets on Chinese Pharmacopoeia
The relevant regulations of reconstituted hepatitis B vaccine titer.Show recombination hepatitis B epidemic disease in the immunogenicity of this antigen mixture and matched group simultaneously
The immunogenicity of Seedling is suitable, and experimental result is shown in Table 3.
Table 3 is containing ridge ash, the immunogenicity experiments result (seropositive conversion of the different components of HIB conjugate, hepatitis B antigen
Rate %)
Note: in the immunogenicity of antigen, serological conversion rate is the meansigma methods of each component.
Compositions 1:HIB GL-PP conjugate+ridge ash antigen stock
Compositions 2:HIB GL-PP conjugate+ridge ash antigen stock+adjuvant
Compositions 1:HIB GL-PP conjugate+ridge ash antigen stock+hepatitis B antigen stock solution
Compositions 2:HIB GL-PP conjugate+ridge ash antigen stock+hepatitis B antigen stock solution+adjuvant
Although, used general explanation, detailed description of the invention and test, the present invention made detailed retouching
Stating, but on the basis of the present invention, can make some modifications or improvements it, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Scope.
Claims (10)
1. an immunogenic composition, it is characterised in that it contains: the Polio virus antigens of inactivation and b type influenza
Haemophilus GL-PP conjugate.
Immunogenic composition the most according to claim 1, it is characterised in that the poliovirus of described inactivation resists
Originally it was sabin I type, sabin II type and sabin type III.
Immunogenic composition the most according to claim 1, it is characterised in that described immunogenic composition contains inactivation
Poliovirus sabin I type antigen 2.5-20D unit/person-portion, sabin II type antigen 5-40D unit/person-portion,
Sabin type III antigen 5-70D unit/person-portion, b type hemophilus influenza GL-PP conjugate 1-15 μ g/ person-portion.
Immunogenic composition the most according to claim 3, it is characterised in that described immunogenic composition contains inactivation
Poliovirus sabin I type antigen 5-15D unit/person-portion, sabin II type antigen 1 0-30D unit/person-portion,
Sabin type III antigen 1 0-40D unit/person-portion, b type hemophilus influenza GL-PP conjugate 5-10 μ g/ person-portion.
Immunogenic composition the most according to claim 1, it is characterised in that described immunogenic composition is possibly together with second
Liver antigen.
6. according to the arbitrary described immunogenic composition of claim 1-5, it is characterised in that described immunogenic composition is also
Including physiologically acceptable carrier.
Immunogenic composition the most according to claim 6, it is characterised in that described immunogenic composition is possibly together with one
Planting adjuvant, described adjuvant is aluminium salt.
Immunogenic composition the most according to claim 1, it is characterised in that the dosage form of described immunogenic composition is
Spray, injection, lyophilized preparation, capsule, tablet or pill.
9. the preparation method of an immunogenic composition, it is characterised in that described method comprises the steps:
(1) the Polio virus antigens stock solution of preparation inactivation；
(2) b type hemophilus influenza GL-PP conjugate is prepared；
(3) the hepatitis B antigen stock solution of preparation inactivation；
(4) the Polio virus antigens stock solution of inactivation is resisted with b type hemophilus influenza GL-PP conjugate and hepatitis B
Stock solution carries out mix and blend, and controls the pH value of mixed liquor between 6-7；
The Polio virus antigens stock solution of described inactivation contains sabin I type antigen 2.5-20D unit/person-portion, sabin
II type antigen 5-40D unit/person-portion, sabin type III antigen 5-70D unit/person-portion, b type hemophilus influenza GL-PP
Conjugate 1-15 μ g/ person-portion；Hepatitis B antigen 5-40 μ g/ person-portion.
10. immunogenic composition described in any one of claim 1-8 is used for preventing or treat poliomyelitis or brain in preparation
Application in film inflammation medicine.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021219047A1 (en) * | 2020-05-01 | 2021-11-04 | 神州细胞工程有限公司 | Method for improving immunogenicity of protein/peptide antigen |
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