CN107184973A - A kind of compound vaccine adjuvant and its application - Google Patents
A kind of compound vaccine adjuvant and its application Download PDFInfo
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Abstract
The present invention provides a kind of compound vaccine adjuvant and its application.Wherein compound vaccine adjuvant is made up of following ingredients：Oil in water emulsion, TLR stimulants and stabilizer.The present invention also provides a kind of immune composition, includes above-mentioned compound vaccine adjuvant and antigen.The compound vaccine adjuvant and antigen combined application, can effectively strengthen the humoral immune reaction and cell immune response of vaccine, its enhancing effect is significantly better than single emulsion, TLR stimulants and aluminum hydroxide adjuvant.The present invention compound vaccine adjuvant cost is low, stability and security are good, can as a variety of vaccines such as Human-papilloma Vaccine, rabies vaccine candidate's adjuvant.
Description
Technical field
The present invention relates to immunological technique field, and in particular to a kind of compound vaccine adjuvant and its in immunization therapy and prevention
The purposes in field.
Background technology
Adjuvant is nonspecific immunomodulator, and adjuvant is added in vaccine, can play raising immune response intensity, change
Become a variety of effects such as immune response type, extension immune response duration.Particularly the vaccine of less immunogenic (is such as gone out
Live vaccine, subunit vaccine, recombinant protein vaccine, polypeptide vaccine etc.), it is immune in difference that rational application adjuvant can improve vaccine
The Conversion rate being inoculated with background crowd.In addition, addition adjuvant can also reduce the consumption of antigen in vaccine, vaccine pin is reduced, is had
Production capacity when helping reduce vaccine cost and solve the problems, such as Epidemic outbreak of disease.
The adjuvant research of early stage is based primarily upon experience, and due to lacking the support of technology and immunologic mechanism research, adjuvant is ground
Hair relatively lags behind in vaccine research and development.From aluminium adjuvant first Application in human body, there is the time of nearly 70 years, it is unique people's adjuvant
(the Jan of O'Hagan DT et al. .Expert Rev Vaccines 2013；12(1):13-30).Although aluminium adjuvant cost is low, peace
Good perfection, but such adjuvant mainly activates the reaction of Th2 para-immunities, does not have invigoration effect to cellular immunity, and in many people's epidemic diseases
There is no the effect of obvious reinforced immunological reaction in seedling (especially recombinant protein vaccine and polypeptide vaccine).Therefore, with more next
Emerging in large numbers for more candidate vaccines and highlighting for vaccine economic benefit, the demand to clinically available novel adjuvant are quickly increasing
It is long.
Emulsion-type adjuvant (including oil in water emulsion, water-in-oil emulsion etc.) is an important branch of novel adjuvant, animal
Experiment display, emulsion can combine a variety of poor antigens and use (recombinant protein, polypeptide etc.), and it is anti-to induce the antigentic specificity of high titre
Body.Oil-phase component, water-phase component and emulsifying agent are three big key elements of emulsion, and the selection of these three key elements and proportioning determine emulsion
Security, stability and immunostimulatory activity.Water-in-oil emulsion is based on oil-phase component, and viscosity is big after emulsification, expands in vivo
Dissipate relatively slow, it is strong to the slow releasing function of antigen, but local irritation and side reaction are larger, therefore limit its application in human body.
And oil in water emulsion, based on water-phase component, human tolerance is higher, and preferable with the compatibility of most vaccine antigens.Due to ore deposit
The nonmetabolizable oil component such as thing oil can cause Regional Body to damage, and cause the toxicity of injection site, therefore people helps
Agent generally selects metabolizable vegetable oil, animal oil etc. (such as peanut oil, squalene) when researching and developing.The emulsifying agent commonly used in emulsion
The mostly good nonionic emulsifier of security.The selection of emulsifying agent is generally according to its hydrophilic lipophilic balance (HLB value), typically
For, HLB value is lower to represent that emulsifying agent hydrophily is weaker, and lipophile is stronger, it is intended to form w/o type emulsion；Conversely, HLB
Value is higher, illustrates that the hydrophily of emulsifying agent is stronger, lipophile is weaker, it is intended to form O/W type emulsion.It is worth noting that,
HLB value can only determine formed emulsion type, but due to the difference of the other specifications such as emulsifying agent structure, molecular size and consumption
Different, the emulsifier performance of identical HLB value is also not quite similar.Research finds, the emulsifying agents of different HLB values is used in mixed way can be with
Stable interface is formed, particle agglutination (Boyd J et al. .J Colloid Interafee Sci, 1972,41 is prevented:359-
370), therefore, often matched in vaccine adjuvant using the emulsifying agent of two kinds and the above.Different oil-phase component, water-phase component
And after emulsifying agent mixing, the interaction mode complexity of various composition is various, therefore, the emulsion prepared using different formulations, its
Adjuvanticity is difficult to predict.
The oil in water emulsion adjuvant of current granted listing has 3 kinds, is MF59, AS03 and AF03 respectively, comprising oil phase
Composition is squalene.Wherein, the granted earliest, applications of MF59 are most wide.MF59 emulsifying agent is Tween-80 and Span-
85, it is the degradable surfactant of human body, is mixed with squalene and buffer solution, the fully emulsified rear oil-in-water emulsion formed
In include diameter about 160nm drop (Jan of O'Hagan DT et al. .Expert Rev Vaccines 2013；12(1):13-
30).Using MF59 as the influenza vaccines of adjuvantIn 1997 in Italy's listing granted first, the current vaccine exists
At least 29 national registrations, cumulative sale amount is more than 60,000,000 doses.During the extensive prevalences of H1N1 in 2009, containing MF59 adjuvant
Two kinds of H1N1 vaccines (With) the also granted crowd (including pregnant woman) for being applied to more than 6 months, it tires out
Meter sales volume has exceeded 100,000,000 doses.In addition, MF59 is used for a variety of viral epidemic diseases such as HIV, CMV, HBV, HCV, HSV also in clinical test
Seedling, and show good security.Studies have shown that MF59 is unable to direct activation DC, but can stimulating expression of macrophage, grain it is thin
Born of the same parents, monocyte secrete cytokines, promote monocyte to be divided into DC, and promote APC to antigen by promoting phagocytosis
Intake；In addition, MF59 can also stimulate the myocyte at immune position, change its gene expression profile, activated immune related gene
(the Jul 18 of Calabro S et al. .Vaccine 2013；31(33):3363-9).MF59 can not activate TLR and Nlrp3 inflammatories
Corpusculum, not yet finds its acceptor directly acted at present, but the performance of its clear and definite adjuvanticity is that MyD88 is relied on
(the Jul 5 of Seubert A et al. .Proc Natl Acad Sci USA 2011；108(27):11169-74).
In addition to emulsion-type adjuvant, the molecule with immunostimulatory activity is also one of developing direction of novel adjuvant.
The TLR stimulants such as particularly Pam3Cys, poly IC, MPL, imiquimod (Imiquimod), CpG ODN, combine a variety of vaccines
Using entering the clinical test (Apr 12 of Steinhagen F et al. .Vaccine 2011；29(17):3341-55).It is this kind of
The leading immune response of the usual activating Th 1 of adjuvant.Wherein Poly IC are artificial synthesized double-stranded RNAs, can be activated simultaneously after being immunized
Humoral immunity and cell immune response, its known receptor are TLR3 and MDA-5 (Yoshida I et al. .Microbiol
Immunol 2013 May；57(5):329-33).TLR3 exempts from the immunocytes such as mDC, T cell and fibroblast etc. are non-
Epidemic disease cell has expression, is the unique TLR independent of MyD88 being currently known, and I types IFN can be caused to secrete after activation, so as to be situated between
Lead DC cell co-stimulatory molecules and MHC II developed by molecule up-regulation (O'Neill LA et al. .Nat Rev Immunol 2007
May；7(5):353-64).MDA-5 can recognize that the dsRNA in cytoplasm, and after poly IC are combined with MDA-5, downstream passes through IPS-
1/FADD/TBK1 paths can also stimulate I types IFN release.Because poly IC have very strong rush cell immune response effect,
It has good application prospect in antiviral and antineoplaston field, at present as glioblastoma, cutaneum carcinoma, ovary
The therapeutic vaccine adjuvant of the tumours such as cancer enters the clinical test (Feb of Ammi R et al. .Pharmacol Ther 2015；146:
120-31).In China, the Polyinosinic injection using poly IC as main component has been used clinically for treatment herpes zoster etc.
Virus infection relevant disease.In addition, the rabies vacciness of adjuvant containing PIKA (main component is poly IC) is carried out in Singapore
Phase I clinical trial, as a result shows that the adjuvant has good security, and the vaccine immunity activity of the adjuvant containing PIKA is better than nothing
Control vaccine (the Jan of Wijaya L et al. .Vaccine 2017 of adjuvant；35:1175–1183).
It is living because the mechanism that the emulsion-type adjuvant being currently known reacts with TLR stimulants activated immune has notable difference
The immune response type of change is also differed, and the two is used in combination, it is expected to which by synergy, more fully activated immune is anti-
Should.The good emulsion of the security TLR stimulant strong with immunostimulatory activity is used in mixed way, is also expected to reduce the consumption of the latter,
By the strength control of immune response within the specific limits, compromise between security and validity.But the adjuvant mixing application of different mechanisms,
Whether there is complementarity, which kind of change can occur for its security and adjuvanticity, and various adjuvant components can with which kind of proportions
Immunostimulatory activity is maximized on the premise of security is ensured, these problems are unpredictable, it is necessary to experimental verification.Therefore,
Inventor presses oil in water emulsion of the present invention and TLR stimulants, particularly MF59 and poly IC and various stabilizers
After certain proportion mixing, with different types of vaccine combined immunization, it is found that two kinds of adjuvant has complementary advantages, not only increase
The antibody level of antigentic specificity, has also effectively activated specific cell immune response, and shown in zoopery
Good tolerance.Therefore, the new adjuvant obtained using this strategy has good application prospect, it is expected to be applied to simultaneously
Preventative vaccine and therapeutic vaccine field.
The content of the invention
It is an object of the invention to provide a kind of new security and immunostimulatory activity be good, answering with human body application prospect
Combination adjuvant, and the immune composition comprising the composite adjuvant, and the composite adjuvant and immune composition are studied in immunization therapy
And the purposes of prevention area.
The present inventor through research surprisingly it has been found that, using appropriate compatibility mode by oil in water emulsion, TLR stimulants
And stabilizer is combined, the composite adjuvant of composition has good security and stronger immunostimulatory activity, and and HPV
VLP vaccines, HPV L2 polyepitope vaccines, HIV are fitted together to various forms of vaccine joints such as VLP vaccines and inactivation rabies vaccine should
With being remarkably improved the immunogenicity of vaccine.The present invention has found based on more than, has completed, and is provided in embodiment hereof
Data.
First aspect present invention provides a kind of adjunvant composition, and it includes oil in water emulsion, TLR stimulants and stabilizer.
Specifically, oil in water emulsion accounts for the 50%-95% of adjunvant composition cumulative volume, the concentration of TLR stimulants is 5 μ g/ml-1000 μ
The tolerable stabilizer concentration of g/ml, human body is 1 μ g/ml-10mg/ml.
Specifically, in the adjunvant composition that first aspect present invention is provided, its oil in water emulsion composition it is main by aqueous phase into
Divide, oil-phase component and emulsifying agent are constituted, the component is appropriately emulsified after mixing in proportion through the mode such as ultrasonic or high-pressure homogeneous, can
Form the emulsifiable suspension of particle stable homogeneous.
It is preferred that, in adjunvant composition of the present invention, the water-phase component that its oil in water emulsion is included is resistant to for human body
The buffer system received, is made up of pH buffer and ionic strength adjustor, and its pH buffer is selected from, but not limited to, ammonium salt, citric acid
Salt, acetate, boric acid, phosphate, sulfate, Tris, butanedioic acid, maleic acid, various amino acid etc.；Its ionic strength adjustor
It is selected from, but not limited to, sodium chloride, potassium chloride, sodium phosphate, potassium phosphate etc..It is particularly preferred, adjunvant composition of the present invention
In, the water-phase component of its oil in water emulsion is made up of phosphate buffer (PBS).The pH value control of water-phase component is resistant in human body
By in the range of, it is preferred that its scope is particularly preferred to be pH 6.5- in pH 5.0-8.5, more preferably pH 6.0-8.0
7.5。
It is preferred that, in adjunvant composition of the present invention, its oil-phase component is metabolizable oil, is selected from, but not limited to, spiny dogfish
Alkene, cholesterol, vegetable oil etc.；Particularly preferred, its oil-phase component of composite adjuvant of the present invention is squalene.Squalene contains
0.25%-10% (v/v) of the amount no more than 10% (v/v), preferably emulsion cumulative volume of emulsion cumulative volume, or 0.5%-
5% (v/v), or 1%-5% (v/v), or 3%-5% (v/v), or 0.5%-1.5% (v/v), particularly preferred, its content is
5% (v/v).
Optionally, in oil in water emulsion of the present invention, various liposoluble constituents can be also included, mother is selected from, but not limited to,
Educating phenol (such as alpha-tocopherol and its derivative), phytosterin compound, phosphatide (such as lecithin, cephalin, cuorin).
It is preferred that, in adjunvant composition of the present invention, the emulsifying agent be surfactant composition, be selected from but
It is not limited to Pluronic L121, mannitol monooleate, polyoxyethylene sorbitan monooleate (Tween 80/polysorbate
80/Tween-80), polyoxyethylene sorbitan laurate (polysorbas20/Tween-20), SPAN85 (this
Disk 85/Span-85), sorbitan fatty ester (span 80/Span-80), triethylene glycol list lauryl ether (Bu Lijie 30/
Brij 30), polyethylene glycol cetyl ether (Bu Lijie 56/Brij56), Triton X-100 (Triton X-100)
Deng；It is furthermore preferred that the emulsifying agent is Tween-80 and Span-85 composition, and the emulsifier content accounts for emulsion totality
Long-pending 0.01%-10% (v/v)；It is preferred that, the content of the emulsifying agent accounts for the 0.3%-3% (v/v) of emulsion cumulative volume；Especially
It is preferred that, Tween-80 content and Span-85 content are 0.5% (v/v) of emulsion cumulative volume in the emulsifying agent.
Particularly preferred, in adjunvant composition of the present invention, the oil in water emulsion is MF59 adjuvant.
The method for preparing oil in water emulsion is that well known to a person skilled in the art (such as patent CN 101522218B).Tool
Body, after the oil-phase component, water-phase component and emulsion are mixed, by repeatedly high-pressure homogeneous, until obtaining comprising required straight
The uniform preparation of footpath oil droplet.In oil in water emulsion of the present invention, comprising oil droplet there is the diameter of submicron order, it is averaged
Diameter is less than 1 μm；It is preferred that, droplet diameter is less than 500nm；It is furthermore preferred that less than 300nm；It is particularly preferred, its diameter range
Between 140-170nm, or between 100-140nm, or between 50-100nm.
In addition, the adjunvant composition described in first aspect present invention, wherein TLR stimulants be selected from TLR1, TLR2, TLR3,
The TLR such as TLR4, TLR5, TLR6, TLR7, TLR8, TLR9 natural or artificial synthesized stimulant, specifically, the TLR is stimulated
Agent is selected from, but not limited to, Pam2CSK4, polyinosinic acid-polycytidylicacid (poly IC), LPS derivatives, flagellin and its mutation
Body, imidazole quinoline and its derivative, CpG etc.；It is preferred that, the TLR stimulants are TLR3 stimulants；It is furthermore preferred that the TLR
Stimulant is oligonucleotide composition；Particularly preferred, the TLR stimulants are poly IC.Poly IC of the present invention
For heterogeneous polynucleotide mixture, its molecular weight is between 66,000 to 1200,000 dalton, the poly preferably
IC molecular weight is particularly preferred between 66,000 to 990,000 dalton, and its molecular weight is in 66,000 to 660,000 road
Between you pause, in a related embodiment, the poly IC used are respectively provided with the molecular weight.
Adjunvant composition described in first aspect present invention also includes the tolerable stabilizer of human body, and the stabilizer has
Combined with TLR stimulants, reduce its toxicity and extend the effect of its half-life period, therefore the immunostimulation of adjuvant can be further enhanced
Activity.
It is preferred that, the stabilizer comprising be selected from, but not limited to, with calcium, cadmium, lithium, magnesium, cerium, caesium, cadmium, cobalt, deuterium, gallium,
The compound of the cations such as iodine, iron or zinc and the cationic compound in inorganic salts or organic double compound form；More preferably
, the stabilizer be selected from, but not limited to, the calcium such as calcium chloride, calcium carbonate, calcirm-fluoride, calcium phosphate, calcium sulfate or calcium gluconae from
Sub- compound；Particularly preferred, the stabilizer includes calcium chloride.
Optionally, stabilizer of the present invention also include the tolerable antibiotic of human body, such as, but not limited to safe hundred mycin,
Anthracycline, butirosin sulfate, gentamicin, hygromycin, amikacin, dibekacin, nebramycin, U.S.
The antibiotic such as its acid amides, neomycin, puromycin, streptomysin or streptozotocin；It is preferred that, stabilizer of the present invention includes ammonia
Base class antibiotic；It is furthermore preferred that stabilizer of the present invention includes kanamycins.
It is particularly preferred, in the adjunvant composition described in first aspect present invention, comprising TLR stimulants and human body be resistant to
The composition for the stabilizer received is PIKA adjuvants, specifically, its composition includes poly IC, calcium chloride and kanamycins.
Optionally, stabilizer of the present invention also includes non-antibiotic class polyamino polyol, these polyaminos
Polyol should have good human tolerance, will not increase adverse reaction when being used in combination with vaccine and adjuvant；It is excellent
Choosing, polyamino polyol of the present invention be the compound that the metabolizable compound of human body or human body can be synthesized and
Its precursor, such as, but not limited to water soluble chitosan, chitosan oligomer, N-Acetyl-D-glucosamine, glycosaminoglycan (mucopolysaccharide) etc..
It is particularly preferred, in the adjunvant composition described in first aspect present invention, comprising TLR stimulants and human body be resistant to
The composition components for the stabilizer received include poly IC, calcium chloride and chitosan oligomer.
It is particularly preferred, in the adjunvant composition described in first aspect present invention, comprising TLR stimulants and human body be resistant to
The composition components for the stabilizer received include poly IC, calcium chloride and N-Acetyl-D-glucosamine.
It is particularly preferred, in the adjunvant composition described in first aspect present invention, comprising TLR stimulants and human body be resistant to
The composition components for the stabilizer received include poly IC, calcium chloride and hyaluronic acid.
It is particularly preferred, in the adjunvant composition described in first aspect present invention, comprising polyamino polyol
Content should can combine most TLR stimulants poly IC, in adjunvant composition, poly IC and polyamino polyhydroxy
The mass ratio of compound is between 1:10 to 10:Between 1；It is preferred that, its mass ratio is between 1:4 to 4:Between 1；It is particularly preferred,
Its mass ratio is between 1:2 to 2:Between 1.
Particularly preferred, in the adjunvant composition described in first aspect present invention, the concentration of calcium chloride is between 0.1mmol/L
To between 100mmol/L.
In adjunvant composition described in first aspect present invention, its oil-phase component has the composition limited with TLR stimulants
Ratio, to ensure the security and immunostimulatory activity of adjunvant composition.In different embodiments, in adjunvant composition
Squalene and poly IC mass ratio are between 1:10 to 15:Between；It is preferred that, its mass ratio is between 1:5 to 12:Between 1；It is special
Not preferred, its mass ratio is between 2:1 to 5:Between 1.It is preferred that, wherein poly IC concentration is between 5 μ g/ml to 1000 μ g/
Between ml.
In a particular embodiment, squalene and poly IC mass ratio can be 0.23:1、0.47:1、2.33:1、4.65:
1、5.81:1 or 11.63:1, different quality than adjunvant composition reinforcing humoral immune reaction and cell immune response ability
There is some difference, but is remarkably improved the immune response level of vaccine induction.
Another aspect of the present invention provides a kind of immune composition, and its composition is as the adjuvant combination described in first aspect present invention
Thing and various forms of antigens composition.Wherein, the antigen may be selected from but be not limited to from bacterium, virus, parasite, true
The antigen of bacterium, tumour, human body autoantigen and/or allergen etc., and the quantity of antigen can be in the immune composition
It is one or more.
It is preferred that, the antigenic source of the immune composition may be selected from the antigen of pathogenic microorganism but be not limited to adenopathy
Malicious (adeniviridae), arenavirus (arenaviridae), astrovirus (astroviridae), Bunyavirus
(bunyaviridae), calicivirus (cliciviridae), flavivirus (flaviviridae), hepatitis viruse
(hepeviridae), Mononegavirales virus (mononegavirales), nest viral (nidovirales), tiny RNA disease
Malicious (piconaviridae), positive myxovirus (orthomyxoviridae), papillomavirus (papillomaviridae),
Parvovirus (parvoviridae), polyomavirus (polyomaviridae), poxvirus (poxviridae), reovirus
(reoviridae), retroviruse (retroviridae), togavirus (togaviridae), actinomyces
(actinobacteria), Chlamydia (chlymidae), sclerine bacterium (firmicutes), mycetozoan (proteobacteria),
The sufficient flagellum door of conveyor screw (spirochaetes), sac fungus (ascomycota), basidiomycetes (basidiomycota), meat
Door (phylum apicomplexa), Ciliophora (phylum are covered in (phylum sarcomastigophora), top
Ciliophora), Platyhelminthes (phylum platyhelminthes), Nematoda (phylum nematode) and/or section
The antigen of main drive thing door (phylum arthropoda) etc..
It is furthermore preferred that the antigen of the immune composition is viral antigen, it may be selected from but be not limited to HPV
(human papillomavirus, HPV), rabies viruses (rabies virus), hepatitis type B virus (Hepatitis B
Virus), influenza virus (influenza virus), poliovirus (poliovirus), human immunodeficiency virus
The multi-form antigen of the virus such as (human immunodeficiency virus, HIV).
Particularly preferred, the antigen of the immune composition is inactivation of viruses, such as, but not limited to：Inactivated rabies virus,
Inactivating influenza virus, inactivated poliovirus, hepatitis A virus, japanese encephalitis virus etc..
Particularly preferred, the antigen of the immune composition is the recombinant protein antigen of viral source, such as, but not limited to：
The virus-like particle of the virus such as HPV (HPV), hepatitis type B virus (HBV), adeno-associated virus (AAV), bacteriophage
(VLP) HPV major cat protein (L1) and/or secondary shell and/or Hybrid virus like particles (cVLP), are included
Recombinant protein antigen of albumen (L2) neutralizing epitope etc..
Especially, as described in embodiment of the present invention, polyinosinic acid-polycytidylicacid and stabilizer calcium chloride and Ka Na are mould
The composite adjuvant that plain (PIKA) joint MF59 adjuvant is constituted, being used in combination with HPV VLP vaccines or inactivation rabies vaccine can have
Effect improves the immunocompetence of vaccine.
Especially, as described in embodiment of the present invention, polyinosinic acid-polycytidylicacid and stabilizer calcium chloride and Ka Na are mould
The composite adjuvant that plain (PIKA) joint MF59 adjuvant is constituted, VLP vaccines chimeric with HIV/HPV, which are used in combination, can effectively improve vaccine
Immunocompetence.
Especially, as described in embodiment of the present invention, polyinosinic acid-polycytidylicacid, calcium chloride, chitosan oligomer or N-
The composite adjuvant that acetylglucosamine joint MF59 adjuvant is constituted, being used in combination with HPV VLP vaccines can effectively improve
The immunocompetence of vaccine.
The invention further relates to the purposes of the adjunvant composition and immune composition, it can be applied to prepare immunization therapy and pre-
The preparation of the multiple use such as anti-, includes but is not limited to：(1) prevention cause pathogeny imcrobe infection and the related various diseases of infection, such as
The infection of influenza virus, rabies viruses, HIV, HPV etc. and infection relevant disease；(2) prevention and treatment of tumour, (3) allergy are anti-
The treatment of immune correlated disease should be waited.
According to the present invention, described adjunvant composition and immune composition can use a variety of formulations, and wherein emulsion components can
Prepared by the way of independent packing or mix with antigen and TLR stimulants is dispensed；When emulsion is using individually packing form, antigen
And/or TLR stimulants can be solution form or lyophilized form.
According to the present invention, described adjunvant composition and immune composition can be connect using the acceptable form of human body
Kind, inoculation position includes but is not limited to parenteral administration, intramuscular injection, intraperitoneal injection, intravenous injection, hypodermic injection, warp
It is respiratory tract suction, rectally, vagina administration, nose administration, oral administration, administration through eye, local administration, transdermal or intracutaneous give
Medicine etc..
The explanation of relational language and explanation
According to the present invention, term " adjuvant " refers to clinically can be applied to human body or having with human body application prospect
Strengthen the material of immune response function, including the various adjuvants that may be got the Green Light with future currently got the Green Light, for example
But it is not limited to aluminium adjuvant, MF59 and various forms of adjunvant compositions.
According to the present invention, term " emulsion " refers to be mixed by proper proportion by water-phase component, oil-phase component and emulsifying agent, warp
The heterogeneous liquid dispersion formed after emulsification.Wherein water-phase component includes but is not limited to phosphate buffer, Tris bufferings
The buffer systems such as liquid, citrate buffer, HEPES buffer solution；Oil-phase component is metabolizable lipid, including but not limited to plant
Oil, fish oil, animal oil, artificial oil and other lipid components (such as, but not limited to squalene, peanut oil, linseed oil, phosphatide etc.)；
Emulsifying agent is the surfactant with suitable HLB, such as, but not limited to SPAN85 (Span-85), poly- sorb
Ester 80 (Tween-80), sorbitan fatty ester (span 80/Span-80), triethylene glycol list lauryl ether (Bu Lijie
30/Brij 30), polyethylene glycol cetyl ether (Bu Lijie 56/Brij 56), Triton X-100 (Triton X-
100) etc..
According to the present invention, term " metabolizable oil " is the oily substance for referring to be converted by organism metabolism, the oil
Can be any vegetable oil, animal oil or artificial oil, they are nontoxic to receiving immune, can be converted by metabolism.
Nut, seed and cereal are common vegetable oil sources；Artificial oil is also the part of the present invention, including can commercialized supply
Various oils.Present invention particularly provides oil-phase component be squalene, be the intermediate of Biosynthesis of cholesterol, be also that one kind can
Metabolism oil.
According to the present invention, term " v/v " refers to the volume/volume ratio of solution each component, can be by following substitution ratio by v/v
The concentration that the concentration conversion of expression is represented into w/v (mass/volume ratio)：5% (v/v) squalene concentration is equal to 4.3% (w/v)
Squalene concentration；0.5% (v/v) Tween-80 concentration is equal to 0.53% (w/v) Tween-80 concentration.
According to the present invention, term " dalton " is international atomic mass unit, can be by following ratio by dalton
The nucleic acid molecular weight of expression is converted into the nucleic acid molecular weight that bp is represented：Length is equal to 660 dongles for 1bp nucleic acid molecular weight
.
According to the present invention, term " stabilizer " refers to be combined and played the composition of stabilization with adjuvant component, including
But it is not limited to antibiotic (such as, but not limited to kanamycins, neomycin, gentamicin), polyamino polyol (for example
But be not limited to chitosan, mucopolysaccharide etc.), inorganic salts (such as, but not limited to calcium chloride, magnesium chloride, calcium phosphate), cation have
Machine compound (such as, but not limited to calcium stearate, calcium gluconae).
According to the present invention, term " antigen " refers to the various materials recognized under appropriate situation by immune system, can come
Come from pathogen, human body itself, tumour etc..
Brief description of the drawings
Fig. 1 shows the poly IC used in composite adjuvant of the embodiment of the present invention relative molecular weight.
Fig. 2 shows in the embodiment of the present invention 2 that the serological specificity induced is immunized in composite adjuvant joint inactivation rabies vaccine
IgG levels.As a result show, the composite adjuvant group of various dose is remarkably improved the specific IgG level of vaccine induction.***：
Compared with without adjuvant group, P<0.001；#：Compared with simple PIKA groups, P<0.05；##：Compared with simple PIKA groups, P<
0.01；###：Compared with simple PIKA groups, P<0.001.
Fig. 3 A to Fig. 3 B are shown in the embodiment of the present invention 3, and mad dog is inactivated using ELISPOT methods detection composite adjuvant joint
The specific cell immunoreaction level that vaccine immunity induces, as a result shows, the composite adjuvant of various dose is remarkably improved
Vaccine induces the level of cell immune response.*：P<0.05；**：P<0.01；***：P<0.001.
Fig. 3 A show the spleens cell number of each group secretion of gamma-IFN；Fig. 3 B show that each group secretes IL-4 spleens cell number.
Fig. 4 shows in the embodiment of the present invention 4 that the Serotypes that composite adjuvant joint HPV VLP vaccine immunities induce are special
Different in nature neutralizing antibody level.**：Compared with without adjuvant group, with P<0.01；***：Compared with without adjuvant group, with P<0.001；#：With
Simple PIKA groups are compared, with P<0.05；###：Compared with simple PIKA groups, with P<0.001；&&&：Compared with simple Alum groups,
With P<0.001.
Fig. 5 shows in the embodiment of the present invention 5 that composite adjuvant combines the serum HPV16 that HPV16L2 multi-epitope antigens induce
Type neutralizing antibody level.***：P<0.001；ND：It is not detected by neutralizing antibody.
Fig. 6 shows in the embodiment of the present invention 6 that composite adjuvant combines the serum HPV16 that HPV16L2 multi-epitope antigens induce
Type neutralizing antibody level.*：Compared with without adjuvant group, P<0.05；**：Compared with without adjuvant group, P<0.01；***：With without adjuvant
Group is compared, P<0.001；###：Compared with simple poly IC groups, with P<0.001；&&&：Compared with simple MF59 groups, with P<
0.001。
Embodiment
The present invention will be further illustrated by non-limiting example below, it is as well known to those skilled in the art, without departing substantially from
In the case of spirit of the invention, many modifications can be made to the present invention, such modification also falls into the scope of the present invention.Below
Embodiment be merely to illustrate the present invention, and should not be taken as limiting the scope of the invention, because embodiment is necessarily various.
The term that is used in this specification is merely to illustrate specific embodiment, rather than as limitation, the scope of the present invention boundary
Determine in the appended claims.
Unless stated otherwise, all technologies as used in this specification and scientific words be and this case art
The meaning that generally understands of technical staff it is identical.The preferred method of the present invention and material are described below, but with this
Any method similar or equivalent with material of method described in specification and material can be used to implement or test the present invention.It is following
Experimental method is conventional method or the method described by product description, used experiment material is such as unless otherwise instructed
Without special instruction, easily it can be obtained from commercial company.All open source literatures being previously mentioned in this specification are incorporated in
This is as reference, to disclose and illustrate the method and/or material in the open source literature.
Embodiment 1：The stability experiment that poly IC are combined with different stabilizers
Poly IC and the composition of different stabilizers are prepared according to the combination shown in table 1, and uses AAS
Poly IC content is determined, 4 DEG C preserved after a period of time, determine residue poly IC content and calculate poly IC preservation
Degree (remaining poly IC contents/initial poly IC contents), it is as a result as shown in table 1 below：
Table 1：Poly IC and different stabilizers combination
*：When calculating poly IC/ stabilizers quality ratios, the quality of calcium chloride is disregarded.
The above results show, poly IC can be effectively stablized using chitosan oligomer or N-Acetyl-D-glucosamine, reduce drop
Solution, can be maintained 12-18 months with the adjunvant composition stability postponed.
Embodiment 2：Composite adjuvant joint inactivation rabies vaccine is immunized mouse and induces antigentic specificity IgG detections
The BALB/c mouse of 4-6 week old is taken, random packet, respectively with inactivation rabies vaccine (RV, Changchun Wei Ersai biological medicaments
Industry Co., Ltd), (solution is MF59 by 5% (v/v) squalene, 0.5% (v/v) Tween-80,0.5%Span-85 (v/v)
PBS) joint RV, polyinosinic acid-polycytidylicacid parenteral solution (PIKA, southern part of the country medicine company) joint RV and MF59/PIKA joints RV are immunized
Mouse, specific packet is as shown in table 2.The volume ratio of MF59 and vaccine other compositions is 1：1.Intramuscular injection, RV immunizing dose
For 0.02IU, it was immunized in the 0th, 2 week, totally 2 times.2nd immune rear 2 weeks tail vein blood, separates serum.
Table 2：Composite adjuvant joint RV experiment mices packet and immunizing dose
(being used for serum IgG detection)
Using inactivation rabies vaccine stoste coated elisa plate (100ng/ holes), after 4 DEG C are incubated overnight, 5%BSA room temperatures are used
Close 2h.Doubling dilution is carried out to mouse immune serum with PBS, is added in the ELISA Plate after closing (100 μ l/ holes), room temperature is incubated
Educate 2h.Using PBS-T (in PBS add 0.05%Tween 20) board-washing 3 times, add PBS-T dilutions HRP- goats resist it is small
Mouse IgG secondary antibodies (1：5000), 37 DEG C of incubation 45min.Using PBS-T board-washings 5 times, developed the color with OPD substrates (Sigma), 37 DEG C incubate
Educate after 5min, use 2M H2SO4Terminating reaction.The OD490 in each hole is read using ELIASA, reading is more than 0.2 and more than the moon
Property 2 times of highest dilution of control be serum RV specific IgG antibodies titres.As a result as shown in Fig. 2 be used in combination MF59 and
The each group antibody level of serum of PIKA adjuvants is all remarkably higher than no adjuvant group and PIKA adjuvant groups is used alone, and MF59/PIKA-
The specific IgG level highest of 10 immune groups.Show the mass ratio when poly IC/ squalenes 0.47:1 to 4.65:Between 1
When, the composite adjuvant that the PIKA adjuvants by MF59 oil in water emulsion and by main component of poly IC are constituted is remarkably improved epidemic disease
Seedling induces the level (using SPSS softwares, One-way ANOVA analyses) of humoral immune reaction, and in the dosage of the present embodiment
Under, do not observe that adjuvant causes toxic reaction in Mice Body.
Embodiment 3：Composite adjuvant joint inactivation rabies vaccine is immunized mouse and induces antigen-specific cellular immune response inspection
Survey
The BALB/c mouse of 4-6 week old is taken, random packet, every group 5, respectively with inactivation rabies vaccine (RV, Changchun Wei Er
Match Bioceuticals Inc.), MF59 (5% squalene, 0.5%Tween-80,0.5%Span-85, buffer solution are PBS) connection
Close RV, polyinosinic acid-polycytidylicacid parenteral solution (PIKA, southern part of the country medicine company) joint RV and MF59/PIKA joints RV and mouse be immunized,
Specific packet is as shown in table 3.The volume ratio of MF59 and vaccine other compositions is 1：1.Intramuscular injection, RV immunizing dose is
0.02IU, it is immune in the 0th, 2 week, totally 2 times.
Table 3：Composite adjuvant joint RV experiment mices packet and immunizing dose
(being used for ELISPOT detections)
2nd time it is immune after take mouse boosting cell within 13 days, antigentic specificity, secretion are detected using ELISPOT methods respectively
The splenocyte of IFN-γ and IL-4.Stimulator antigen is the RV vaccinogen liquids of inactivation, and concentration is 4 μ g/ml.As a result such as Fig. 3 A and Fig. 3 B
Shown, MF59/PIKA-10 groups secretion of gamma-IFN and IL-4 cell number are significantly higher than other each groups, and MF59/PIKA-5 components
The cell number for secreting IFN-γ and IL-4 is significantly higher than no adjuvant group, nonantigenic group and independent MF59 adjuvant group, but less than MF59/
PIKA-10 groups.Show the mass ratio when poly IC/ squalenes 2.33:1 to 4.65:When between 1, by MF59 oil in water emulsion
And the composite adjuvant that the PIKA adjuvants by main component of poly IC are constituted is remarkably improved vaccine induction cell immune response
Level (uses SPSS softwares, One-way ANOVA analyses), and under the dosage of the present embodiment, does not observe adjuvant in mouse
Cause toxic reaction in vivo.The method of ELISPOT detections is disclosed, such as patent CN 101166559B.
Embodiment 4：Composite adjuvant joint HPV VLP vaccine immune mouses induce the detection of neutralizing antibody level
The BALB/c mouse of 4-6 week old is taken, random packet (is obtained from Chinese medicine section with HPV16L1 Δs N5C31VLP respectively
Institute of Basic Medical Sciences of institute Xu Xuemei laboratories) combine polyinosinic acid-polycytidylicacid parenteral solution (PIKA, southern part of the country medicine company) and hydrogen
Aluminum adjuvant (Alum) or MF59 adjuvant (5% squalene, 0.5%Tween-80,0.5%Span-85, buffer solution are PBS)
Immune mouse, specific packet and immunizing dose are as shown in table 4.The volume ratio of MF59 and vaccine other compositions is 1：1.Subcutaneous note
Penetrate, it is immune in the 0th, 2 week, totally 2 times.2nd immune rear 2 weeks tail vein blood, separates serum.
Table 4：Composite adjuvant joint HPV VLP vaccine immunities experiment mice packet and dosage
The HPV16 neutralizing antibody titers of immune serum are detected using HPV16 pseudovirus, as a result as shown in figure 4, single
The neutralizing antibody level solely induced using aluminum hydroxide adjuvant or PIKA adjuvants HPV16L1VLP is made without being remarkably reinforced effect
Combine the neutralizing antibody level that PIKA adjuvants are remarkably improved VLP inductions with MF59, and with exclusive use aluminium hydroxide or individually
Compared using PIKA adjuvants, neutralizing antibody level is also significantly improved.Show the mass ratio when poly IC/ squalenes 0.23:
1 to 11.63:When between 1, the compound assistant that the PIKA adjuvants by MF59 oil in water emulsion and by main component of poly IC are constituted
Agent is remarkably improved the immunocompetence (using SPSS softwares, One-way ANOVA analyses) of vaccine, and in the dosage of the present embodiment
Under, do not observe that adjuvant causes toxic reaction in Mice Body, with application prospect.Pseudovirus prepare and pseudovirus in and test
Method be disclosed, such as patent CN 104418942A.
Embodiment 5：The detection that mouse induces neutralizing antibody level is immunized in composite adjuvant joint HPV16L2 polyepitope vaccines
The hFc γ RI transgenic mices (C57BL/6 backgrounds) of 4-6 week old are taken, HPV16L2 multilists are used in random packet respectively
Position antigen (containing 3 L2aa.17-36 epitopes copied and hFc γ RI targetings domain) (is obtained from Chinese Academy of Medical Sciences basis doctor
Learn research institute and permitted snow plum laboratory) combine polyinosinic acid-polycytidylicacid parenteral solution (PIKA, southern part of the country medicine company), MF59 adjuvant (5%
Squalene, 0.5%Tween-80,0.5%Span-85) immune mouse, specific packet and immunizing dose are as shown in table 5.MF59 with
The volume ratio of vaccine other compositions is 1：1.It is subcutaneously injected, it is immune in the 0th, 2,4,6 week, totally 4 times.2 weeks tails are quiet after the 4th is immune
Arteries and veins is taken a blood sample, and separates serum.
Table 5：Composite adjuvant combines HPV16L2 multi-epitope antigen immunization experiment mice groups and dosage
The HPV16 neutralizing antibody titers of immune serum are detected using HPV16 pseudovirus, as a result as shown in figure 5, making
With MF59 combine PIKA adjuvants be remarkably improved HPV16L2 multi-epitope antigens induce neutralizing antibody level, and with exclusive use
MF59 or exclusive use PIKA adjuvants are compared, and neutralizing antibody level is also significantly improved.Show the matter when poly IC/ squalenes
Amount is than being 11.63:When 1, the composite adjuvant that the PIKA adjuvants by MF59 oil in water emulsion and by main component of poly IC are constituted
The immunocompetence (using SPSS softwares, One-way ANOVA analyses) of vaccine is remarkably improved, and in the dosage of the present embodiment
Under, do not observe that adjuvant causes toxic reaction in Mice Body, with application prospect.Pseudovirus prepare and pseudovirus in and test
Method be disclosed, such as patent CN 104418942A.
Embodiment 6：Composite adjuvant joint HPV16L2 polyepitope vaccines containing different stabilizers, which are immunized mouse and induce to neutralize, to be resisted
The detection of body level
The hFc γ RI transgenic mices (C57BL/6 backgrounds) of 4-6 week old are taken, HPV16L2 multilists are used in random packet respectively
Position antigen (is obtained from Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences to be permitted to avenge plum laboratory) (the L2aa.17-36 tables containing 3 copies
Position and hFc γ RI targetings domain) combine the immune mouse of the composite adjuvant containing different stabilizers, specific packet and immunizing dose are such as
Shown in table 6.The volume ratio of MF59 and vaccine other compositions is 1：1.It is subcutaneously injected, it is immune in the 0th, 2,4,6 week, totally 4 times.4th
Secondary immune rear 2 weeks tail vein bloods, separate serum.
Table 6：Composite adjuvant combines HPV16L2 multi-epitope antigen immunization experiment mice groups and dosage
The HPV16 neutralizing antibody titers of immune serum are detected using HPV16 pseudovirus, as a result as shown in fig. 6, making
The neutralizing antibody level that HPV16L2 multi-epitope antigens induce is remarkably improved with the composite adjuvant of 4 kinds of formulas, and with individually making
Compared with MF59 or exclusive use poly IC adjuvants, neutralizing antibody level is also significantly improved.Show by MF59 oil-in-waters breast
The immunocompetence that the composite adjuvant that agent, poly IC and polyamino polyhydroxy based stabilizer are constituted is remarkably improved vaccine (uses SPSS
Software, One-way ANOVA analyses), and under the dosage of the present embodiment, do not observe that adjuvant causes toxicity anti-in Mice Body
Should, with application prospect.Method in pseudovirus preparation and pseudovirus with experiment is disclosed, such as patent
CN104418942A。
Embodiment 7：Composite adjuvant joint HPV/HIV is fitted together to VLP vaccine immune mouses and induces AntiHIV1 RT activity neutralizing antibody level
Detection
The BALB/c mouse of 4-6 week old is taken, random packet is fitted together in HIV and table with 2 kinds by skeleton of HPV16VLP respectively
Chimeric VLP (being permitted snow plum laboratory obtained from Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences) connection of position (2F5 neutralizing antibodies epitope)
Close composite adjuvant and mouse is immunized, specific packet and immunizing dose are as shown in table 7.The volume ratio of MF59 and vaccine other compositions is 1：
1.It is subcutaneously injected, it is immune in the 0th, 2,4,8 week, totally 4 times.2 weeks tail vein bloods after the 4th is immune, separate serum.
Table 7：Composite adjuvant joint HPV/HIV is fitted together to VLP immunization experiments mice group and dosage
The HIV neutralizing antibodies of immune serum are detected using HIV sensitive strain SF162 pseudovirus, as a result such as the institute of table 8
Show, composite adjuvant is remarkably improved the neutralizing antibody for HIV epitopes that chimeric VLP vaccines induce.
Table 8：Composite adjuvant joint HPV/HIV is fitted together to the immune HIV neutralizing antibodies level detections induced of VLP
Claims (10)
1. a kind of adjunvant composition, it includes oil in water emulsion, TLR stimulants and the tolerable stabilizer of human body, its reclaimed water bag
Oil emu accounts for the 50%-95% of adjunvant composition cumulative volume, the concentration of TLR stimulants is 5 μ g/ml-1000 μ g/ml, human body can
The stabilizer concentration of tolerance is 1 μ g/ml-10mg/ml.
2. the adjunvant composition according to claim 1, wherein oil in water emulsion include oil-phase component, water-phase component and breast
Agent；
The oil-phase component includes squalene, and its content is the 1%-10% (v/v) of oil in water emulsion cumulative volume, it is preferred that its
Content is 3%-5% (v/v), it is furthermore preferred that its content is 5% (v/v)；
The water-phase component includes the tolerable pH buffer of human body and ionic strength adjustor, wherein the human body is tolerable
PH buffer includes citrate or phosphate, and the ionic strength adjustor includes sodium chloride and/or potassium chloride, the aqueous phase
The pH value of composition in the range of 6.0-8.0,
It is preferred that, the water-phase component is PBS；
The emulsifying agent is the composition of nonionic surface active agent, it is preferred that the emulsifying agent is Tween-80 and Span-
85 composition；
It is further preferred that Tween-80 content and Span-85 content are respectively that oil in water emulsion is total in the emulsifying agent
The 0.3%-3% (v/v) of volume；
Particularly preferred, Tween-80 content and Span-85 content are oil in water emulsion cumulative volume in the emulsifying agent
0.5% (v/v)；
Particularly preferred, the oil in water emulsion is MF59 adjuvant.
3. adjunvant composition according to claim 1 or 2, wherein TLR stimulants are natural TLR stimulants or artificial conjunction
Into TLR stimulants, wherein the TLR include but is not limited to TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8,
TLR9；
It is preferred that, the TLR stimulants are TLR3 stimulants；
It is furthermore preferred that the TLR stimulants are oligonucleotide composition；
It is particularly preferred, the TLR stimulants be polyinosinic acid-polycytidylicacid poly IC, its molecular weight between 66,000 to
Between 1200,000 dalton, particularly between 66,000 to 660,000 dalton.
4. the tolerable stabilizer of adjunvant composition according to any one of claim 1 to 3, wherein human body include with
TLR stimulants combine, reduce its toxicity and extend the compound of its half-life period；
It is preferred that, the tolerable stabilizer of human body includes the compound with cation, and/or the tolerable antibiosis of human body
Element, and/or non-antibiotic class polyamino polyol, wherein,
The cation is selected from calcium, cadmium, lithium, magnesium, cerium, caesium, cadmium, cobalt, deuterium, gallium, iodine, iron or zinc, the change with cation
Compound is in the form of inorganic salts or organic double compound；
It is furthermore preferred that the tolerable stabilizer of human body is more comprising ionic calcium cpd, and/or antibiotic, and/or polyamino
Hydroxy compounds, wherein,
The ionic calcium cpd is selected from calcium chloride, calcium carbonate, calcirm-fluoride, calcium phosphate, calcium sulfate or calcium gluconae,
It is mould that the antibiotic is selected from safe hundred mycin, anthracycline, butirosin sulfate, gentamicin, hygromycin, amikacin
Element, dibekacin, nebramycin, its beautiful acid amides, neomycin, puromycin, streptomysin or streptozotocin,
The polyamino polyol is selected from water soluble chitosan, chitosan oligomer, N-Acetyl-D-glucosamine, osamine and gathered
Sugar；
It is particularly preferred, the tolerable stabilizer of human body comprising calcium chloride and/or kanamycins, and/or chitosan oligomer,
And/or N-Acetyl-D-glucosamine, and/or hyaluronic acid.
5. adjunvant composition according to any one of claim 1 to 4, wherein TLR stimulants and human body are tolerable steady
It is PIKA adjuvants to determine agent, and it includes poly IC, calcium chloride and the card molecular weight between 66,000 to 660,000 dalton
That mycin.
6. adjunvant composition according to any one of claim 1 to 4, wherein,
The TLR stimulants are poly IC of the molecular weight between 66,000 to 660,000 dalton,
The tolerable stabilizer of human body is calcium chloride and polyamino polyol, the polyamino polyol
Selected from chitosan oligomer, N-Acetyl-D-glucosamine and hyaluronic acid,
The mass ratio of poly IC and polyamino polyol is between 1:10 to 10:Between 1, the concentration of calcium chloride between
Between 0.1mmol/L to 100mmol/L.
7. the adjunvant composition according to claim 1 to 6, wherein poly IC concentration are between 5 μ g/ml to 1000 μ g/ml
Between, and poly IC and squalene mass ratio between 1:10 to 15:Between 1；It is preferred that, its mass ratio is between 1:5 to 12:1
Between；Particularly preferred, its mass ratio is between 2:1 to 5:Between 1.
8. a kind of immune composition, it includes adjunvant composition as claimed in any of claims 1 to 7 in one of claims and antigen,
Optional, the antigen derives from bacterium, virus, parasite, fungi, tumour, human body autoantigen to be one or more
And/or the antigen of allergen；
It is preferred that, the antigenic source is in adenovirus adeniviridae, arenavirus arenaviridae, astrovirus
Astroviridae, Bunyavirus bunyaviridae, calicivirus cliciviridae, flavivirus flaviviridae, liver
Scorching virus hepeviridae, Mononegavirales virus mononegavirales, nest virus nidovirales, tiny RNA disease
Malicious piconaviridae, positive myxovirus orthomyxoviridae, papillomavirus papillomaviridae, tiny disease
Malicious parvoviridae, polyomavirus polyomaviridae, poxvirus poxviridae, reovirus reoviridae,
Retroviruse retroviridae, togavirus togaviridae, actinomyces actinobacteria, Chlamydia
Chlymidae, sclerine bacterium firmicutes, mycetozoan proteobacteria, conveyor screw spirochaetes, sac fungus
Door is covered on the sufficient flagellum door phylum sarcomastigophora of ascomycota, basidiomycetes basidiomycota, meat, top
Phylum apicomplexa, Ciliophora phylum ciliophora, Platyhelminthes phylum platyhelminthes,
Nematoda phylum nematode and/or Arthropoda phylum arthropoda；
It is furthermore preferred that the antigenic source is in HPV, rabies viruses, hepatitis type B virus, influenza virus, spinal cord ash
The scorching virus of matter, human immunodeficiency virus.
9. immune composition according to claim 8, wherein the antigen is inactivation of viruses, the inactivation of viruses, which is selected from, to go out
Rabies viruses living, influenza virus, poliovirus, hepatitis A virus, japanese encephalitis virus.
10. immune composition according to claim 8, wherein the antigen for viral source recombinant protein antigen and/
Or polypeptide or epitope antigen, it is selected from：HPV or the virus-like particle VLP of hepatitis type B virus, embedded virus sample
Grain cVLP, HPV major cat protein L1 and/or the recombinant protein antigen or polypeptide in secondary coat protein L2 sources
Or epitope antigen.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2023165573A1 (en) * | 2022-03-02 | 2023-09-07 | 北京市希波生物医学技术有限责任公司 | Culture medium formula for activating whole anti-tumor immune system and method for preparing agonist activated whole immune effector cells |
| CN117100852A (en) * | 2023-10-24 | 2023-11-24 | 江苏瑞科生物技术股份有限公司 | Composite adjuvant and preparation method and application thereof |
| CN118059227A (en) * | 2024-04-22 | 2024-05-24 | 江苏瑞科生物技术股份有限公司 | Application of oil-in-water emulsion in preparation of rabies immune suit |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2571925A (en) * | 2018-03-08 | 2019-09-18 | Univ Antwerpen | Iron-based vaccine adjuvants |
| JP2021529787A (en) * | 2018-06-29 | 2021-11-04 | 信福（北京）医▲薬▼科技有限公司 | How to prepare a complex to enhance the immune response |
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| WO2023165573A1 (en) * | 2022-03-02 | 2023-09-07 | 北京市希波生物医学技术有限责任公司 | Culture medium formula for activating whole anti-tumor immune system and method for preparing agonist activated whole immune effector cells |
| CN115645523A (en) * | 2022-12-22 | 2023-01-31 | 深圳大学总医院 | Application of polymer lipid hybrid nanoparticles as immunologic adjuvant and immunologic preparation |
| CN115645523B (en) * | 2022-12-22 | 2023-03-21 | 深圳大学总医院 | Application of polymer lipid hybrid nanoparticles as immunologic adjuvant and immunologic preparation |
| CN117100852A (en) * | 2023-10-24 | 2023-11-24 | 江苏瑞科生物技术股份有限公司 | Composite adjuvant and preparation method and application thereof |
| CN118059227A (en) * | 2024-04-22 | 2024-05-24 | 江苏瑞科生物技术股份有限公司 | Application of oil-in-water emulsion in preparation of rabies immune suit |
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| Publication number | Publication date |
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| CN107188932A (en) | 2017-09-22 |
| CN107188932B (en) | 2020-02-11 |
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