CN104922684A

CN104922684A - Naturally-sourced and/or self-assembled biological membrane, a closed structure or cell compartment with biological membrane property and preparation method and application thereof

Info

Publication number: CN104922684A
Application number: CN201510282308.4A
Authority: CN
Inventors: 高颖; 姚震祥; 魏建良; 王旻子
Original assignee: HANGZHOU YOUMADA BIOTECHNOLOGY Co Ltd
Current assignee: HANGZHOU YOUMADA BIOTECHNOLOGY Co Ltd
Priority date: 2015-05-28
Filing date: 2015-05-28
Publication date: 2015-09-23

Abstract

The invention relates to a naturally-sourced and/or self-assembled biological membrane and a closed structure and a cell compartment with the biological membrane property and a preparation method and an application thereof. The biological source of the biological membrane, the closed structure or the cell compartment with the biological membrane property is animals, plants or microorganisms, an obtained biological membrane is complete or fragmental and of a lipoid two-molecular layer structure on the morphology, constituents of the biological membrane are lipoid and proteins, and little sugar is combined to the lipoid or proteins by virtue of a covalent bond. The biological membrane, the closed structure of the cell compartment with the biological membrane property can be applied to a biological medical carrier technology and particularly can be applied to a transgene carrier and drug carrier; the biological membrane, the closed structure of the cell compartment with the biological membrane property can be used for researching and developing product of cosmetics additives and cosmetics active ingredient carriers, also can be used for researching and developing products of vaccines and immunomodulators and also can be used for researching and developing products on the aspect of high polymer materials.

Description

The biomembrane that natural origin and/or self-assembling technique obtain, there is the closing structure or cellular compartment and its preparation method and application of biomembrane

Technical field

The invention belongs to biomedicine and polymeric material field, be specifically related to the biomembrane obtained by natural origin and/or self assembly and the closing structure and cellular compartment and its preparation method and application with biomembrane.

Background technology

Biomembrane (biomembranes) or biological film (biological membranes) are the general names of all membrane structures of cell, organelle and its environment junction, playing a part to divide and separate cell and crganelle, is also the significant points relevant with intracellular communication.In biology except some virus, all there is biomembrane.Eukaryotic cell, except plasma membrane (also known as cell membrane), separates the membranous system of various organelle in addition, comprises nuclear membrane, mitochondrial membrane, endoplasmic reticulum, lysosome membrane, Golgi membrane, chloroplast membranes, vacuole, peroxisome film etc.

Biomembrane form is all the lamellar structure of lipid bilayer (lipid bilayer), its constituent mainly lipid, separately has a small amount of protein and sugar class.Biomembranous this lipid bilayer Rotating fields, makes it have following functions: the turnover of regulation and control material, the packaging of material and transport between different organelle in cell, to provide specific trafficking pathways and provide the space of material storage by forming intracellular compartment (cellular compartment) for some reagent and semiochemicals.

Biomembranous component is various, the difficulty of separation and purification is larger, therefore in one's early years for ease of research, often adopt the various synthetic membrane structures of single one or several lipid composition: unimolecular layer membrane, built up film, liposome, dull and stereotyped bilayer lipid membranes etc., in addition, also after protein can being embedded, composition rebuilds film, these membrane structure general terms " synthetic membrane ".Synthetic membrane is applied in practice, as from the solution such as sea water mutually in efficiently isolation and identification material, as the dialyzer of Patients With Kidney Diseases and for clinical diagnosis and treatment etc., utilized liposome can prepare with features such as cell membrane fusion another larger expansion that liposome vectors medicine is synthetic membrane in recent years.

But, comprise these synthetic membranees of liposome, all have in vitro oxidizable, seepage, storage characteristics are bad; Easily can not be arrived shortcoming such as target tissue performance useful effect etc. by the degraded of some enzyme materials and macrophage phagocytic in vivo, these all limit its application as carrier.On the other hand, the interpolation of synthetic material, makes synthetic membrane inevitably make body produce rejection as macromolecular material implant into body, uses limited.

Have in living matter and its long-term evolution process from simple to complex on earth, biomembranous appearance once leaps, biomembrane just has fine structure so and exquisite functional activity mechanism today through the evolution of 1 years, itself there are two maximum characteristics, i.e. the mobility of film and membrane assymmetry.The mobility of film refers to that biomembrane is in continuous kinestate, and this makes the continuous switch of the lipid molecular in film, and it ensures that film has the essential condition of the functions such as endocytosis, exocytosis, material Transfer, cell fusion.Exactly because but also this membrane fluidity causes the continuous distortion of film, make component relative to factors such as the instability of the better simply synthetic membrane morphosis of natural biological film, cause liposome interior synthetic membrane as vector stabilisation and storage characteristics poor, wrappage easily leaks.In addition, biomembrane can be divided into inside and outside nearly kytoplasm face and non-kytoplasm face two-layer, inside and outside biomembrane, the component of two layers and structure have very big-difference, this species diversity is called biomembranous unsymmetry, this membrane assymmetry produces important impact to the sequence of film, cell fusion, intermolecular identification just, and this unsymmetry is synthetic membrane can not have completely.

On the other hand, life is the highest form that material exists, and the most basic feature that life has is: the teaching display stand regulation and control by metabolism, self replication and oneself's assembling.The constitution element that self assembly (self-assembly) is system, refers to not by under the intervention of mankind's external force, self-assemble, is organized into the phenomenon of regular texture.Self assembly is the basis that various complex biological structure is formed, inseparable with biosis, and biomembrane is the natural model of the self assembly research that organism provides to people.

Summary of the invention

In order to overcome the weak point of the synthetic membrane remained at present at applied defect, an object of the present invention is to provide biomembrane, the closing structure with biomembrane or cellular compartment that natural origin and/or self-assembling technique obtain; Another object of the present invention is to provide biomembrane that natural origin and/or self-assembling technique obtain, has the closing structure of biomembrane or the preparation method of cellular compartment; 3rd object of the present invention is to provide above-mentioned biomembrane, has the application of the closing structure of biomembrane or cellular compartment.This biomembrane, the closing structure with biomembrane or cellular compartment can be applicable to biological medicine carrier technique, in particular for transgene carrier, pharmaceutical carrier; Research and the product development of cosmetics additive and cosmetic industry Ingredients Carrier can be applied to; Also can be applicable to research and the product development of vaccine and immunomodulator; Also research and the product development of the aspects such as macromolecular material can be applied to.

In order to realize first above-mentioned object, present invention employs following technical scheme:

The biomembrane that natural origin and/or self-assembling technique obtain, the closing structure with biomembrane or cellular compartment, the biogenetic derivation of this biomembrane, the closing structure with biomembrane or cellular compartment is animal, plant or microorganism, the biomembrane obtained is complete or fragment, for being lipid bilayer Rotating fields in form, its constituent is lipid and protein mainly, separately has a small amount of saccharide by being covalently bonded on lipid or protein.

As preferably, described biomembrane, the closing structure with biomembrane or cellular compartment, particle diameter is from 10nm to tens microns.

As preferably, described biomembrane, the shape of closing structure or cellular compartment with biomembrane comprise the variform structure of spherical, vesicle shape, shaft-like, spiral helicine monolayer or multilamellar, multicell.

As preferably, described biomembrane comprise plasma membrane, nuclear membrane, mitochondrial membrane, endoplasmic reticulum, lysosome membrane, Golgi membrane, chloroplast membranes and vacuole and peroxisome film one or more.

As preferably, described cellular compartment refers to general organelle; As preferred again, described cellular compartment be mitochondrion, chloroplast, peroxisome, lysosome, endoplasmic reticulum, nucleus, Golgi body and vesicle and microtubule one or more.

In order to realize second above-mentioned object, present invention employs following technical scheme:

Biomembrane described in an above-mentioned arbitrary technical scheme, have the closing structure of biomembrane or a preparation method for cellular compartment, the method comprises the following steps:

1) biological cell is obtained;

2) by the mass propgation under its adapt circumstance of the cell described in step 1);

3) obtain step 2) described in the lysate of cell, and by differential centrifugation, density－gradient centrifuga－tion method and two-phase extraction method, various method is used alone or combination of two or triple combination use and extract required biomembrane, the closing structure with biomembrane and cellular compartment.

As preferably, described differential centrifugation extracting method comprises the following steps:

1. by cell pyrolysis liquid with 15,000 ~ 30,000 × g, 10 ~ 30min, high speed centrifugation under 1 ~ 6 DEG C of condition, abandons precipitation, collect supernatant;

2. supernatant is with 100, and 000 ~ 200,000 × g, 30 ~ 90min, ultracentrifugation under 1 ~ 6 DEG C of condition, abandons supernatant, collecting precipitation, is the required biomembrane, the closing structure with biomembrane or the cellular compartment that extract, the PBS/ normal saline resuspended preservation of precipitation containing 15 ~ 30% glycerol.

As preferably, described density gradient centrifugation extracting method comprises the following steps:

1. by Eddy diffusion after lysis liquid precipitate, re-suspension liquid joins in the sucrose solution of variable concentrations, with 150, and 000 ~ 300,000 × g, 60 ~ 90min, ultracentrifugation under 1 ~ 6 DEG C of condition, collects required layer;

2. the liquid collected with 100,000 ~ 200,000 × g, 30 ~ 90min, ultracentrifugation under 1 ~ 6 DEG C of condition, abandons supernatant, collecting precipitation, is the required biomembrane, the closing structure with biomembrane or the cellular compartment that extract, the PBS/ normal saline resuspended preservation of precipitation containing 15 ~ 30% glycerol.

As preferred again, the mass percent concentration scope of described step 1. sucrose solution is 10 ~ 70%; As preferably, the different quality percent concentration of described step 1. sucrose solution is 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%.

As preferably, described two-phase extracting process comprises following feature:

1. configure the water biphase mixture of glucosan/Polyethylene Glycol, mix homogeneously in separatory funnel, 4 DEG C of standing stay-over demixions, careful separation levels, makes fresh upper and lower phase;

2. Eddy diffusion after lysis liquid precipitate is joined in water biphase mixture, put upside down 30 ~ 40 mix homogeneously up and down gently;

3. with 2,000 ~ 4.000 × g, 5 ~ 10min, centrifugal under 4 DEG C of conditions, get upper and lower phase and continue to enter binary system, be separated three times and merge upper phase afterwards, after diluting 5 times, 60,000 ~ 100.000 × g, 30 ~ 90min, centrifugal under 4 DEG C of conditions, collecting precipitation, is the required biomembrane, the closing structure with biomembrane or the cellular compartment that extract, the PBS/ normal saline resuspended preservation of precipitation containing 15 ~ 30% glycerol.

As preferred again, described two-phase is the biphase mixture of glucosan/Polyethylene Glycol.

As preferred again, described two-phase comprise water two-phase or, organic biphasic, aqueous phase solution and organic phase solution, described solvent is selected from any one or its combination of water, acetonitrile, acetone, oxolane, methanol, ethanol, propanol.

A kind of self assembly is prepared biomembrane, is had the method for the closing structure of biomembrane or cellular compartment, the method comprises the preparation method described in each technical scheme above-mentioned, and biomembrane step 3) obtained, there is the closing structure of biomembrane and the material of cellular compartment covers on wall of a container with the form of desciccator diaphragm, then slowly water or buffer solution is injected, and slight or violent vibration, biomembrane, the closing structure with biomembrane and cellular compartment needed for self assembly obtains.

As preferably, described step 3) obtained material dissolves in chloroform solvent, join in container, reduction vaporization makes biomembrane sprawl on vessel surface, add PBS buffer solution after being evaporated to constant weight and slowly shake 0.5 ~ 3h, in 100,000 ~ 200,000 × g, 30 ~ 90min, ultracentrifugation under 1 ~ 6 DEG C of condition, abandons supernatant, collecting precipitation, is required biomembrane, has the closing structure of biomembrane and cellular compartment.

In order to realize the 3rd above-mentioned object, present invention employs following technical scheme:

The above-mentioned biomembrane described in any one technical scheme, the closing structure with biomembrane or cellular compartment application, this is applied as described biomembrane, have the closing structure of biomembrane or cellular compartment to wrapping up in the film of active component, in film parcel, surface adsorption, surface-crosslinked, intermembranous inlay and wrap up in film add targeting.

As preferably, described active component comprises vaccine or immunomodulatory activity composition, cosmetics or cosmetic active ingredient, active constituents of medicine, genetic stew and cell or tissue.

Biomembrane of the present invention, the closing structure with biomembrane or cellular compartment can be applicable to biological medicine carrier technique, in particular for research and the product development of transgenic, pharmaceutical carrier and cosmetic industry Ingredients Carrier; Also can be applicable to research and the product development of vaccine and immunomodulator; Research and the product development of the aspect such as immunologic diagnosis, macromolecular material can also be applied to.This invention proposes closed biomembrane that natural origin and/or self-assembling technique obtain is applied to the aspects such as chemical industry, medicine and cosmetics imagination as carrier and/or immunomodulator first, and develops multiple corresponding macromolecular material, medicine and cosmetic material and intermediate.

pharmaceutical carrier:

Biomembrane of the present invention, the closing structure with biomembrane or cellular compartment (hereinafter referred to as biomembrane), as pharmaceutical carrier, can reach the following effect improving medicine:

One, stability is improved: due in biomembrane bimolecular lamellar lipid membrane and intermembranous encapsulating effect, medicine and extraneous unstable factor touch opportunity can be made to reduce, stability raising.Such as benzylpenicillin, unstable to acid, oral easily by stomach acids destroy; After biomembrane encapsulating, then can improve the effect of its stability and oral absorption.

Two, dissolubility is improved: the difficult insoluble chemical molecule of most drug, the solvent adding a lot of toxic side effect is often needed to carry out solubilising during use, and biomembrane is as after carrier entrapped drug, amphiphilic due to its lipid bilayer, can be dissolved into the ingredient of indissoluble in the middle of water soluble medicament or fat-soluble medicament.Such as paclitaxel is the broad-spectrum anti-cancer drug of a line, but the dissolubility in water is very little, so with the addition of a large amount of ethoxylate castor oil solubilisings in the paclitaxel prescription of listing both at home and abroad, but ethoxylate castor oil exists larger sensitization, the problem such as bronchospasm, rapid breathing, tired, hypotension can be caused; If use biomembrane envelope paclitaxel, improving deliquescent while, there is allergy hardly.

Three, improve curative effect: because biomembrane has the feature of cellular affinity and histocompatibility, medicine can fully be permeated to target cell target tissue.As antituberculotics rifampicin is encapsulated in biomembrane, medicine can be loaded in people's cell and kills tulase, curative effect can be improved significantly compared with traditional Rimactane.

Four, reduce toxicity or zest: some chemotherapeutics has very strong vascular toxicity, the vascular complication that chemotherapeutic phlebitis etc. is common can be caused, and medicine carries out the vascular toxicity that intravenous injection can reduce medicine significantly again after being encapsulated by biomembrane.On the other hand, there is larger nephrotoxicity in some medicine especially antibiotics and antitumor class medicine, after medicine is encapsulated by biomembrane, can effectively concentrate in the more rich organs of monokaryon-macrophage such as liver, spleen and bone marrow and make medicine cumulant specific ionization medicine in the heart, kidney low, to the heart, the virose medicine of kidney or to the virose medicine of normal cell through encapsulating after, obviously can reduce the toxicity of medicine.Such as amphotericin B is the classical medicine for systemic fungal infection, but amphotericin B toxicity is larger, especially nephrotoxicity, life-time service can cause kidney and Circulatory involvement, significantly limit its application, and after being encapsulated with biomembrane, its distribution in vivo can be changed, obviously reduce toxicity.

Five, slow release long-acting: many medicines are in vivo due to rapid metabolization or excretion, therefore action time is short.Medicine biomembrane is encapsulated, excretion and metabolism can be reduced, the prolong drug holdup time in blood, make medicine slow releasing in vivo, thus extend the action time of medicine.Such as calcitonin secretes by parafollicular cells of thyroid gland the polypeptide hormone produced, often be used to clinically treat osteoporosis diseases, but the half-life of polypeptide drugs is short, often needs repetitively administered, and by encapsulating calcitonin with biomembrane, its prolonged half-life in vivo one times can be made.

Six, targeting: be divided into passive target and active targeting two kinds.Passive target refers to that the medicine through biomembrane parcel enters in human body and can be engulfed as extraneous foreign body by macrophage, is mainly engulfed by the macrophage of monokaryon-macrophage system institute and absorbs, the passive targeting of reticuloendothelial system such as formation liver, spleen etc.Such as, after protostib (meglumine antimonate) is encapsulated by biomembrane as the medicine for the treatment of liver leishmaniasis, the concentration of medicine in liver improves 500 times.And active targeting refers to link targeting factor as part on the biomembrane enclosing medicine, can the specific receptors bind with target cell, thus change microgranule NATURAL DISTRIBUTION in vivo and arrive specific target site.Such as amycin is broad-spectrum anti-tumor medicine, but it has very strong cardiac toxicity, time serious, even heart failure can be caused, but amycin is by after the biomembrane parcel containing cancer target factor R GD, simultaneously, lung cancer therapy effect at least doubles obvious reduction cardiac toxicity.

transgenic transfection reagent:

Transgenic technology refers to that DNA fragmentation is transferred in particular organisms, recombinates with the genome of itself, then from recombinant, carry out the artificially breeding counting generation, thus obtains the individuality with the specific hereditary character of stable performance.This technology can make restructuring biology increase desired new character on the one hand, cultivates new varieties; On the other hand, Gene transfer techniques is not only the important tool of research transgenic and gene expression, and is the committed step of current gene therapy.Desirable gene transfection agent should have following features: high-efficiency transfection, safety, low cytotoxicity, method be simple, save time, economical.But, now conventional transfection reagent be not transfection efficiency too low be exactly that cytotoxicity is too large, therefore, need badly find a kind of newly can reduce Cytotoxic transfection reagent again by high-efficiency transfection.

Biomembranous lipid bilayer structure makes itself and cell membrane have good similar compatibility, can be adsorbed by cell membrane, again by the fusion of film or the endocytosis of cell, once in a while also by direct osmosis, thus can as allogenic material as DNA enters the carrier of cell.DNA transmits and enters cell, and form inclusion body or enter lysosome, wherein sub-fraction DNA can discharge and enter Cytoplasm in inclusion body, then enters in core further and transcribe, express.Meanwhile, biomembranous natural origin, produces cytotoxicity hardly.

cosmetics:

Horny layer is the outermost layer of skin, and the lipid in horny layer is the intensive duplicature wrapped up, and alternately, the two combines alternately with covalency for it and corneocyte, the cuticular main part of common formation, is also the main matter basis of skin barrier function.Have research display: as long as remove lipid from horny layer of epidermis, skin barrier function just can be made to lose, and moisture content in skin significantly reduces, skin occurs dry; Stop the lipid in removing skin, then in horny layer, moisture is recovered thereupon, and xerosis cutis improves.Illustrate thus, lipid is present between skin keratinocytes, is filled with intercellular substance, plays binding agent effect, stops that in skin, moisture is to external diffusion, keeps moisture, soft skin, isolate from outside skin simultaneously, have barrier and moisture-keeping functions by foreign body.

Biomembrane has lipid bilayer Rotating fields, makes it have following functions when cosmetic field is applied:

One, moisture-keeping function: the lipid components such as the cholesterol in biomembrane, ceramide and Palmic acid, the lipid barrier of skin can be repaired, also obviously can increase skin electric conductivity simultaneously, make it have the ability of very strong associated water molecule, network structure is formed in horny layer, to maintain moisture of skin, improve skin elasticity.

Two, whitening function: the lipid components such as the ceramide in biomembrane, often as signaling molecule, can regulate the peroxidization of intracellular peroxide and enhancing lipid, have whitening effect; On the other hand, containing a large amount of unsaturated fatty acid in biomembrane, the deposition of melanin on skin can be reduced, make whiteness of skin.

Three, anti-aging effects: the phospholipid in biomembrane can enter deep skin, and be combined with the phospholipid of the deep skin cell membrane thing that originates from, and make cell membrane fluidization, as can be increased mobility and the permeability of film containing the unsaturated phospholipid of linoleic acid and alpha-linolenic acid, thus strengthen the metabolic function of cell, play the effect of activating cell.Impaired when running into cell tissue, when disease or aging, reparation and the growth of epidermal growth factor can be promoted, thus reach delaying cell aging.

Being more than biomembrane adds as the functional component of cosmetics, plays the effect of its uniqueness; Biomembrane as the carrier of functional components, can also reach the following effect improving functional components simultaneously:

One, stability is improved: due in biomembrane bimolecular lamellar lipid membrane and intermembranous encapsulating effect, functional components and extraneous unstable factor touch opportunity can be made to reduce, stability raising.Such as, containing more unsaturated double-bond in retinol structure, to light and heat stability poor, be easy to oxidized after ingress of air, after biomembrane encapsulating, its stability strengthens greatly.

Two, dissolubility is improved: the very most of difficult soluble substance of beauty functions composition, be difficult to during use play its efficacy effect through skin barrier, amphiphilic due to lipid bilayer, indissoluble effect of encapsulating can be divided as carrier and penetrated in the middle of water-soluble substances and liposoluble substance by biomembrane.

Three, long-lasting: biomembrane also has slow releasing function as carrier.Experiment proves, biomembrane and the entrapped drug retention time in blood circulation, and it is many that majority wants the retention time of specific ionization medicine to grow.

Four, penetration enhancement: the functional components in cosmetics must be transmitted through horny layer and reaches corresponding site of action, just can play nutrition, improve the effect of skin.And the horny layer of the skin of people has extremely strong barrier action, macromolecular functional components is difficult to penetrating.With biomembrane packaging function composition, because biomembrane is similar to horny layer structure, have good affinity, functional components carries the increase of lower percutaneous transit dose biomembranous.

vaccine carrier and immunomodulator:

The development of the subjects such as biology, molecular immunology and genetic engineering makes vaccine have more and more consequence, and the effective adjuvant of application safety and carrier carry out vaccine delivery by increasing people is familiar with.Complete Freund's adjuvant, incomplete Freund's adjuvant, bacterial endotoxin, polyanion and mineral adsorbent that use is in the past more, they all produce local and systemic toxicity, form unacceptable granuloma, because effect duration is short, effectiveness is low, therefore gradually abandon by people.Aluminium glue adjuvant is safe and effective, but it can only cause humoral immunization, also exist can not inducing cellular immune and antigen in conjunction with shortcomings such as differences between batches are larger.

Therefore obtain a kind of safety, effectively, the vaccine carrier of the efficient inducing cellular immune of energy and humoral immunization and adjuvant new technique have become the new challenge of vaccine application.

Biomembranous lipid bilayer structure makes itself and cell membrane have good similar compatibility, can be adsorbed by cell membrane, again by the fusion of film or the endocytosis of cell, once in a while also by direct osmosis, the carrier of cell thus can be entered as allogenic material.Biomembrane is by wrapping up or the mode such as absorption, and can be used as the carrier of the materials such as albumen, nucleic acid, synthetic peptide, cytokine, antibacterial, virus, meanwhile, biomembrane is a kind of non-pathogenic carrier, and safety coefficient is high.On the other hand, biomembrane can be absorbed by body antigen presenting cell effectively, thus evokes potent immunoreation, and therefore itself has again natural immunoadjuvant function.

Biomembrane, as vaccine carrier and immunomodulator, has following advantage:

One, safety is high: biomembrane is the carrier of a kind of non-immunogenic and non-pathogenic as the antigen presentation of vaccine and delivery vehicle, and safety coefficient is high, and does not cause granuloma at inoculation position, and repeated inoculation is also without any ill effect.Meanwhile, can not detect after biomembrane immune animal that the antibody for biofilm carrier produces, reduce the probability that biofilm carrier itself causes immunity of organism rejection.

Two, multi-functional: the material such as the orientable assembling albumen of biofilm carrier, nucleic acid, synthetic peptide, cytokine, antibacterial, virus, can not only be applicable to the structure of traditional vaccine, also can apply to novel gene vaccine as carrier and adjuvant.

Three, compound property: can the different material such as antigen, nucleic acid of NW-TFT on biomembrane, can develop multivalence type combination vaccine and multiple vaccines.

Four, efficient humoral immunity: dendritic cell (DC cell) is the professional antigen that the function of discovery is at present the strongest is delivery cell (APC), biomembrane is due to its lipid bilayer Rotating fields, the form merged by after birth promotes that APC is to the picked-up of antigen, thus the humoral immunization effectively activated for corresponding antigens produces antibody, a lot of infectious disease of prevention is played a significant role.

Five, strong cellular immunity: traditional vaccine is main mainly with exciting humoral immunization, but HIV, HCV and tumor etc. need can the novel therapeutic type vaccine of inducing specific cellular immunization, this has higher requirement to the carrier of vaccine and adjuvant.Biofilm carrier has natural advantage, effectively can activate DC cell and other antigen presenting cells, and pass through by ectogenic antigen presentation to MHC-1, and activate CD8+ and the CD4+ T lymphocyte for antigen, obtain potent cellular immunization activation.

Six, slow-releasing: biofilm carrier link antigen, can play the effect that antigen delays to discharge, cause more thoroughly immunne response, improve the effect of immunity.

The application of biomembrane vaccine:

One, the preventative vaccine of infectious disease: in the prevention of antibacterial, virus and parasitic infection, humoral immunization has significant effect.By the immunoreation of biofilm carrier link micro-, modified but identical with pathogen antigen transfer body, can prevent, control the generation of infectious disease, popular.

Two, the therapeutic vaccine of infectious disease: for the body of viral infection, the antibody that humoral immunization produces cannot be removed and enter intracellular pathogen, and preventative vaccine just loses meaning.Viral disease there is no any effective medicine so far, but its characteristics of incidence is again based on intracellular infection, therefore can inducing potent specific cell immune response by biofilm carrier vaccine, providing effective approach for solving problems.

Three, the preventative vaccine of tumor: the infection of persistence high-risk human mammilla papillomavirus (HPV) result in the generation of nearly all human invasive cervix neoplasms; The infection of helicobacter pylori (H.pylori) is the important paathogenic factor of the disease such as gastric cancer, mucosa tissue lymphoma (MALT), and World Health Organization (WHO) has been classified as a class cancerigenic factor.Etiology comparatively clear and definite between this cancerigenic factor and tumor contacts, the antigen can be correlated with by biofilm carrier link or gene preparation vaccine, be inoculated in the healthy population or high-risk group with susceptibility, and then the generation of tumor can be controlled, the research and development of tumor prevention vaccine are expected to contain tumor from " source ".

Four, the therapeutic vaccine of tumor: the research of the therapeutic vaccine of tumor at present concentrates on immunization therapy and gene therapy two aspects, organism immune response can be improved in different levels, promote the adjusting function in cellular-restoring its own amplification cycle, to reach the object for the treatment of tumor simultaneously.The focus of research is effectively to improve tumour immunity and break immune tolerance.Prepare peptide vaccine, recombinant vector vaccine, dendritic cell (DC) vaccine etc. by biofilm carrier, stronger specific T-cells reaction can be induced, thus strengthen the inhibitory action to tumor, gross tumor volume is diminished.

Five, autoimmune disease vaccine: the immunologic injury of autoimmune disease can cause corresponding histoorgan generation pathological changes.The therapeutic vaccine of autoimmune disease produces immunoreactive memory t cell to design to autologous tissue's antigen mainly for removal, and it is auxiliary with immunoregulatory method, the cytokine that some has immunoloregulation function as added, makes immune system adjusted on the whole.Prepared the modes such as DNA vaccination, combined vaccine, fused polypeptide vaccine by biofilm carrier, make it have a good application prospect in autoimmune disease vaccine.

macromolecular material:

Since ancient times, the mankind just have such imagination: if disease appears in some organs of health, can change organ as machine renewal part.1954, in physician's Hartwell Harrison of boston, u.s.a and Joseph write from memory, have successfully completed first case human organ transplant operation---kidney transfer operation, start the New Times of human organ transplant since then.But, the development obstacles that organ transplantation runs into also allows doctors' rather headache of countries in the world, topmost obstacle is, when patient carries out organ transplantation, its body is repelled unavoidably, at present, the medicine of the lifelong all adhib Immunosuppressions of patient of organ transplantation is accepted, to prevent from occurring rejection in body, but these medicines, simultaneously again to whole immune system generation effect, can reduce the ability of patient's resist the disease.In order to address this problem, scientist starts to develop a kind of new method: by the local organization of patient self or cell, in conjunction with some macromolecular materials as support, turns out the organ of " intimate " in the region of interest of patient body.This macromolecular material support will have good cell, histocompatibility, for their growths provide place; Require that there is degradation characteristic simultaneously, harmless after degraded, and any sequela is not stayed to tissue and organ.But also lack desirable material in order to manufacture artificial organ support at present.

Using biomembrane as substrate, cell or tissue is cultivated above, as the stent applications of artificial organ or regeneration and restoration product in organ transplantation, not only there is safety high, rejection can not be produced in implant into body, and performance and originally organize very nearly the same, availability is high, and the investigation and application for macromolecular material provides abundant technical support.

Accompanying drawing explanation

Fig. 1 is hbs antigen content.

Fig. 2 is Symwhite-337-biomembrane result of use figure.

Fig. 3 is Transdermal absorption concentration under different time.

Fig. 4 B16 cell suppression ratio.

Detailed description of the invention

Below in conjunction with specific embodiment and accompanying drawing, set forth the present invention further.Should be appreciated that these embodiments only can not be used for for illustration of the present invention limiting the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.

the biomembranous extraction of embodiment 1, purification

Employing density－gradient centrifuga－tion method extracts, purification obtains required biomembrane.Detailed process is as follows:

(1) liver is got in the execution of the new zealand white rabbit of jejunitas 18-24h, after removing large blood vessel, liver is cut into 2mm ³the fritter of left and right, and bleach to piece of tissue with normal saline flushing;

(2) homogenate is prepared: 1mmol/L CaCl ₂, 50mmol/L HEPES (pH 7.4), 1mmol/L PMSF, 2 μ g/mL aprotiniies, 2 μ g/mL Antipain;

(3) by liver organization and homogenate in quality: the ratio of volume adds the homogenate of 8 times of volumes, and in ice bath environment with electric homogenate machine 15,000rpm homogenate to organizing liquefaction;

(4) by homogenate through four layers of filtered through gauze, filtrate 1,000 × g, 10min, centrifugal under 4 DEG C of conditions, collecting precipitation;

(5) precipitation sucrose solution A(0.3mol/L sucrose, 50mmol/L Tris, 3mmol/L MgCl ₂) fully suspend precipitation, adds the sucrose solution B(2mol/L sucrose of 9 times of volumes, 50mmol/L Tris, 3mmol/L MgCl ₂), upper strata sucrose solution C(0.25mol/L sucrose, 10mmol/L HEPES, 1mmol/L EDTA) fill up centrifuge tube, 90,000 × g, 150min, centrifugal under 4 DEG C of conditions, collect upper strata rete;

(6) this rete washing liquid (50mmol/L HEPES, 1mmol/L PMSF, 2 μ g/mL aprotiniies, 2 μ g/mL Antipain) washing, 90,000 × g, 60min, centrifugal under 4 DEG C of conditions, collecting precipitation, is required biomembrane.

the biomembranous extraction of embodiment 2, purification

Employing Aqueous two-phase partition extracts, purification obtains required biomembrane.Detailed process is as follows:

(1) screen full normal corn seed unanimously, through 1%NaClO soaking disinfection 10min, rinse well, accelerating germination 72h under 25 DEG C of constant temperature dark conditions;

(2) Semen Maydis epicotyl is got, with quality: the ratio of volume adds the Extraction buffer of 2 times of volumes (5mmol/L EDTA, 25mmol/L Tris, 0.25mmol/L sucrose, 1mmol/L MgSO ₄, 0.2%(W/V) and BSA, 0.5%(W/V) PVP-10,10%(W/V) glycerol, 15mmol/L beta-mercaptoethanol, 1mmol/L PMSF, 1mmol/L DTT), ice bath is ground to liquefaction;

(3) by lapping liquid through four layers of filtered through gauze, filtrate 12,000 × g, 15min, centrifugal under 4 DEG C of conditions, get supernatant;

(4) supernatant is 80,000 × g, 30min, centrifugal under 4 DEG C of conditions, collecting precipitation;

(5) precipitation suspension (25mmol/L Tris, 0.25mmol/L sucrose, 0.2%(W/V) BSA, 10%(W/V) mannitol, 1mmol/L DTT) fully suspend precipitation;

(6) suspension is added binary system (to contain in every 10g binary system: 1.7g sucrose, 0.003g DTT, 2.25mL water, 50mmol/L KCl 0.5mL, 1.63 PEG, 3.25g Dextran T-500,200mmol/L PBS 0.5mL), shake up 50 times up and down, 4,000 × g, 5min, centrifugal under 4 DEG C of conditions, get upper and lower phase to continue to enter binary system, be separated three times and merge upper phase afterwards, after diluting 5 times, 80,000 × g, 60min are centrifugal under 4 DEG C of conditions, collecting precipitation, is required biomembrane.

the biomembranous extraction of embodiment 3, purification

Employing differential centrifugation extracts, purification obtains required biomembrane.Detailed process is as follows:

(1) Antarctic ice microalgae ( chlamydomonas subcaudata) be separation and purification from Antarctic Sea Ice;

(2) Antarctic ice microalgae is inoculated in culture medium (containing 21.2g NaCl, 3.6g NaSO in every 10L culture medium in the ratio of 1:100 ₄, 0.6g KCl, 0.3g NaHCO ₃, 0.1g KBr, 0.1g H ₃bO ₃, 0.1g NaF, 9.6g MgCl ₂6H ₂o, 1.0g CaCl ₂, 0.1g SrCl ₂6H ₂o), be placed in controlled illumination box to cultivate.At-4 DEG C, light intensity is 1300 ~ 1900lx, periodicity of illumination 12 is bright/and 12 dark, cultivate 14 days under shaking 4 ~ 5 conditions every day;

(3) by ice algae culturing liquid at 4,000rpm, 20min, centrifugal under 4 DEG C of conditions, collect ice algae precipitation, and rinse rapidly 2 times with the distilled water of pre-cooling, to remove the impurity in extracellular dope, salt surfactant and culture fluid;

(4) above-mentioned ice algae of collecting is precipitated in quality: the ratio of volume adds homogenate buffer (the 0.5mol/L KOH of 4 times of volumes, 0.5mol/L sucrose, 3mmol/L EDTA, 0.6% PVP, 1mmol/L PMSF, 1mmol/L DTT, 5mmol/L ascorbic acid, 0.6% BSA), with squash type cell crushing instrument smudge cells;

(5) by the cell homogenates liquid that obtains at 8,000rpm, 20min, centrifugal under 4 DEG C of conditions, collect supernatant;

(6) supernatant is 145,000 × g, 60min, centrifugal under 4 DEG C of conditions, and collecting precipitation, is required biomembrane.

the separation of embodiment 4 cellular compartment, purification

Employing two-phase extraction is separated, purification obtains required cellular compartment.Detailed process is as follows:

(1) preparation of water two-phase system: by mixture (containing 90g 20%(W/W in every 300g) Dextran T-500,45g 40%(W/W) PEG 3350,33.9g sucrose, 7.5g 0.2mmol/L PBS, 0.45g 2mmol/L KCl) mix homogeneously in centrifuge tube, make the water biphase mixture of isoconcentration glucosan/Polyethylene Glycol (Dextran T-500/PEG 3350), mix homogeneously in separatory funnel, 4 DEG C of standing stay-over demixions, careful separation levels, make fresh upper and lower phase, preserve respectively, for following purification steps in 4 DEG C;

(2) biomembrane precipitation resuspension buffer (5mmol/L PBS, 0.33mol/L sucrose, 3mmol/L KCl, 1mmol/L DTT, 1mmol/L PMSF, the 0.1mmol/L EDTA) resuspension will obtained in embodiment 3;

(3) above-mentioned heavy suspension is added in Dextran T-500/PEG 3350 water biphase mixture of preparation in step (1) with the ratio of mass ratio 1:3, puts upside down 30 ~ 40 mix homogeneously up and down gently;

(4) mixed liquor is at 1,500rpm, 10min, centrifugal under 4 DEG C of conditions, get phase liquid and lower phase liquid continues to enter binary system, be separated the liquid that to be separated on merging afterwards for three times, after diluting 5 times, 100,000 × g, 60min, centrifugal under 4 DEG C of conditions, collecting precipitation, is required cellular compartment, and is rich in thylakoid.

the separation of embodiment 5 cellular compartment, purification

Employing density－gradient centrifuga－tion method is separated, purification obtains required cellular compartment.Detailed process is as follows:

(1) 10g robust growth is chosen, the Spinach Leaf preferably grown under continuous several fine day, arteries and veins in removing after cleaning, in quality: the ratio of volume adds Extraction buffer (the 50mmol/L kaliumphosphate buffer of 6 times of volumes, 0.3mmol/L sorbitol, 2mmol/L EDTA, 1mmol/L MgCl ₂, 1mmol/L MnCl ₂, 1% BSA, 1mmol/L DTT), ice bath grinds;

(2) Percoll layering liquid (50mmol/L HEPES-KOH, 0.3mmol/L sorbitol, 2mmol/L EDTA, 1mmol/L MgCl is configured ₂, 1mmol/L MnCl ₂, 1% BSA, 3%PEG 6000,1%Ficoll), 30,000 × g, 20min are centrifugal in advance under 4 DEG C of conditions;

(3) by lapping liquid four layers of filtered through gauze, 3,000 × g, 15min, centrifugal under 4 DEG C of conditions, collecting precipitation, precipitation 2ml Extraction buffer suspends, and is placed on the Percoll layering liquid of step (2) centrifugal mistake, 15,000 × g, 20min, centrifugal under 4 DEG C of conditions, draw lower floor, be required cellular compartment, and be rich in chloroplast.

the biomembranous preparation of embodiment 6

Adopt the biomembrane needed for self-assembling technique preparation.Detailed process is as follows:

(1) biomembrane obtained in above-described embodiment 1 or embodiment 2 or embodiment 3 is dissolved in enough chloroform solvents;

(2) join in round-bottomed flask by above-mentioned biomembrane chloroformic solution, reduction vaporization makes biomembrane sprawl on Flasks surface, adds PBS buffer solution and slowly shake 2h after being evaporated to constant weight;

(3) by above-mentioned solution at 6,000rpm, 20min, centrifugal under 4 DEG C of conditions, collecting precipitation, and with resuspended buffer (20mmol/L Tris, 0.1mol/L NaCl, 2mmol/L MgCl ₂, 1mmol/L DTT) and resuspended precipitation;

(4) by re-suspension liquid 150,000 × g, 60min, ultracentrifugation under 4 DEG C of conditions, abandons supernatant, collecting precipitation, is the biomembrane obtained by self-assembling technique, and has closing characteristics.

parcel in embodiment 7 film

Reverse evaporation is adopted to carry out DNA parcel in biomembrane, specific as follows:

(1) by the biomembrane that obtains in above-described embodiment 1 or embodiment 2 or embodiment 3 and 18-amine. with 10:1(volume ratio) ratio mixing, be dissolved in the chloroform of 4 times of volumes, then reduce pressure rotary evaporation (37 DEG C, 200rpm) on a rotary evaporator, removing chloroform;

(2) plasmid containing hepatitis B virus DNA is dissolved in the PBS of 20mmol/L;

(3) add the biomembrane in ether dissolution step (1), and mix with the above-mentioned PBS equal-volume containing plasmid, be placed in 50 DEG C of water-baths and hatch 20min, be the required transgene carrier having wrapped up hepatitis B virus DNA plasmid.

parcel in embodiment 8 film

Adopt supersound method in biomembrane, carry out the ingredients of cosmetics, specific as follows:

(1) under 4 DEG C of conditions, vitamin C is dissolved in the PBS buffer solution of 1mol/L, forms saturated solution;

(2) the cellular compartment mixing will obtained in said vitamin C saturated solution and embodiment 5, use sonicator ultrasonic emulsification in ice bath environment, the amplitude of ultrasonic probe is 40%, and work 30sec, interval 30sec, circulate 20 times;

(3) by emulsion 100,000 × g, 20min, centrifugal under 4 DEG C of conditions, abandon supernatant, collecting precipitation, be and required wrapped up ascorbic cosmetics carrier.

embodiment 9 surface adsorption

Electrostatic Absorption method is adopted to prepare vaccine at biofilm surface viral adsorption, specific as follows:

(1) using soda acid potentiometric titration to calculate isoelectric point, IP the biomembrane obtained by self-assembling technique in embodiment 6 is 6.8;

(2) above-mentioned biomembrane is redissolved in pH be in the PBS buffer of 5.8, water bath sonicator 10min;

(3) by known, isoelectric point, IP be 4.8 swine fever virus and above-mentioned redissolution fully mix at the biomembrane isoconcentration of PBS, mixing be placed on-30 DEG C of freeze overnight, then-60 DEG C of lyophilized overnight, are required swine Fever Vaccine.

embodiment 10 is surface-crosslinked

Cross-linking method is adopted to prepare artificial organ at biofilm surface crosslinking cell, specific as follows:

(1) biomembrane obtained by self-assembling technique in embodiment 6 is cross-linked through glutaraldehyde solution, prepares biomembrane template;

(2) human meniscus fibrocartilage cells is placed in incubator (5% CO ₂, 37 DEG C) cultivate, get second filial generation cell F-12 culture fluid (containing 10% calf serum) and make cell suspension;

(3) the dry template of biomembrane through ultraviolet radiation sterilization is put into 6 well culture plates, add above-mentioned cell suspension and submergence it, be placed in constant incubator the 2h that vibrates at 37 DEG C after, culture plate taken out and is placed in CO ₂incubator (5% CO ₂, 37 DEG C), cultivate 2 days, be required artificial meniscus.

embodiment 11 is intermembranous inlays

High pressure homogenization method is adopted to inlay medicine between biomembrane bilayer, specific as follows:

(1) by the cellular compartment that obtains in embodiment 4 and retinoic acid in 65 DEG C of meltings, obtain colostrum with high-shearing dispersion emulsifying machine high shear;

(2) colostrum is mixed into 65 DEG C be dissolved with the distilled water of tween 80 after, high pressure homogenize 15 times circulation, homogenization pressure is 80MPa, naturally cools to room temperature, is required to have wrapped up the pharmaceutical carrier of retinoic acid.

in embodiment 12 film, parcel adds targeting

Adopt pH gradient method to carry out medicine parcel in biomembrane, and utilize the albumen link targeting factor on biomembrane, specific as follows:

(1) be dissolved in the aqueous solution of 0.3mol/L citric acid (pH=4) by the biomembrane obtained in above-described embodiment 1 or embodiment 2 or embodiment 3, rotation of then reducing pressure makes biomembrane form thin film on the wall;

(2) regulate biomembrane obtained above with the sodium carbonate liquor of 1mol/L, and make the pH to 7.8 of suspension, make biomembrane inside and outside formation proton gradient, can be used as carrier Active loading;

(3) under 60 DEG C of conditions, amycin is dissolved in the HEPES buffer solution (pH=7.8) of 1mol/L, forms the saturated solution of amycin;

(4) by biomembrane suspension and the mixing of amycin saturated solution, at 60 DEG C of Water Unders bath hatching 20min, by mixed liquor 80,000 × g, 60min, centrifugal under 4 DEG C of conditions, abandon supernatant, collecting precipitation, be and required wrapped up the pharmaceutical carrier of amycin;

(5) with Avidin labelling targeting factor iRGD, and the above-mentioned biofilm carrier being loaded with amycin is modified upper biotin, by biotin-avidin system, iRGD is connected to and is loaded with on the biofilm carrier of amycin, be and required wrapped up the pharmaceutical carrier of amycin, and there is targeting.

embodiment 13 pharmaceutical carrier beneficial effect (improving medicine/cosmetics stability)

(1) the ascorbic carrier 5%Triton X-100 that wrapped up obtained in embodiment 8 is jiggled at 37 DEG C the breakdown of emulsion that spends the night, and with the methanol dilution of volume ratio 100 times;

(2) carry out assay by high performance liquid chromatography to vitamin C, calculating Vitamin C content in carrier is 8.72%;

(3) by 8.72% Vitamin C content vitamin C and carrier simply mixed in contrast with wrapped up ascorbic carrier and be kept in the flint glass bottle of sealing respectively, carry out stability test: 9d(intensity 3000lx is penetrated in illumination), sampling and measuring two groups of ascorbic changes of contents every three days;

(4) result shows, the Vitamin C Stability wrapped up by biomembrane is apparently higher than mixture.

embodiment 14 pharmaceutical carrier beneficial effect (improving medicine/cosmetics dissolubility)

Supersound method is adopted to carry out medicine parcel in biomembrane, specific as follows:

(1) under 4 DEG C of conditions, paclitaxel is dissolved in ethanol, form saturated solution;

(2) the cellular compartment mixing will obtained in above-mentioned paclitaxel saturated solution and embodiment 5, use sonicator ultrasonic emulsification in ice bath environment, the amplitude of ultrasonic probe is 40%, and work 30sec, interval 30sec, circulate 20 times;

(3) by emulsion 100,000 × g, 20min, centrifugal under 4 DEG C of conditions, abandon supernatant, collecting precipitation, be and required wrapped up the pharmaceutical carrier of paclitaxel;

(4) the pharmaceutical carrier PBS having wrapped up paclitaxel by above-mentioned acquisition is resuspended to 1ml, and jiggles at 37 DEG C the breakdown of emulsion that spends the night with 5%Triton X-100, and with the methanol dilution of volume ratio 100 times;

(5) carry out assay by high performance liquid chromatography to paclitaxel, calculating in 1ml solution content of taxol in carrier is 9.1%;

(6) excessive paclitaxel raw material is put in PBS buffer solution, jiggle for 37 DEG C, the changes of contents of sampling and measuring paclitaxel, no longer changes to concentration at set intervals, adopt high performance liquid chromatography, calculate the content of paclitaxel for 6.0ug/mL with external standard counter point;

(7) result shows, the dissolubility of the paclitaxel wrapped up by biomembrane improves 1500 times.

embodiment 15 pharmaceutical carrier beneficial effect (raising curative effect of medication)

Reverse evaporation is adopted to carry out medicine parcel in biomembrane, specific as follows:

(1) biomembrane obtained in above-described embodiment 1 or embodiment 2 or embodiment 3 is dissolved in the chloroform of 4 times of volumes, the rotary evaporation that then reduces pressure on a rotary evaporator (37 DEG C, 200rpm), removing chloroform;

(2) rifampicin is dissolved in PBS, adds the biomembrane in ether dissolution step (1), and mix with above-mentioned rifampicin equal-volume, be placed in 50 DEG C of water-baths and hatch 20min, be and required wrapped up the pharmaceutical carrier of rifampicin;

(3) the pharmaceutical carrier 5%Triton X-100 having wrapped up rifampicin by above-mentioned acquisition jiggles at 37 DEG C the breakdown of emulsion that spends the night, and with the methanol dilution of volume ratio 100 times, carry out assay by high performance liquid chromatography to rifampicin, calculating rifampicin content in carrier is 10.2%;

(4) get healthy mice 20, male and female are not limit, body weight 25-35g, inject 1.75 × 10 from tail vein ⁸the H of colony-forming units (CFU)/mL ₃₇r _vtubercule bacillus suspension 0.1mL makes it infect, and injects latter 14 days, mice is divided into A at random, B two groups, often organizes 10;

(5) intravenous injection in every three days of the pharmaceutical carrier that wrapped up rifampicin of A group by above-mentioned acquisition once, and each 10ul, B group injects identical rifampicin medicine dosage in contrast;

(6) injection is expired by sacrifice after three weeks, and get spleen homogenate and be incubated on the Russell medium inclined-plane of debita spissitudo, after six weeks, calculating clump count also calculates CFU;

(7) result shows, is doubled by the more common rifampicin treatment effect of rifampicin that biomembrane wraps up.

embodiment 16 pharmaceutical carrier beneficial effect (improving curative effect of medication, different dosage form)

(1) get healthy new zealand white rabbit 20, male and female are not limit, and body weight 1.5 ~ 2.0kg, is divided into A at random, B two groups, often organize 10;

(2) with after the pentobarbital sodium auricular vein injecting anesthetic of 3%, on temporo, quadrant cuts off bulbar conjunctiva, row gas urges vitreous body art, and namely after limbus of corneae, 3mm sentences No. 30 syringe needles to vitreous chamber central authorities injection C ₃f ₈gas 0.1ml, makes during gas expansion, vitreous body to be pressed to periphery, after 3d, in kind after anesthesia, sentence 5-0 silk thread at former inserting needle mouth and make the preset suture of a cotton-padded mattress formula, micro-vitreous-body-retina lancination wears sclera, release gas, inject L929 cell suspension, set up PVR experimental model;

(3) inject after cell, the pharmaceutical carrier having wrapped up retinoic acid obtained in embodiment 11 is injected 0.2ml, B group and injects the Retinoic Acid In Silicone Oil medicine 0.2ml that obtains on the market in contrast by A group, the preset suture of ligation;

(4), after vitreous of rabbit eyes injects L929 cell, form mist-like bleary district at posterior vitreous immediately, and gradually became linen band at about one week, connect to nipple and medullary ray, postoperative 28d, after tropicamide mydriasis, indirect ophthalmoscopy vitreous body situation.Vitreous opacity degree is divided into 0 ~ III 4 grade.0 grade: vitreous body is without muddiness, I grade: vitreous body is slightly muddy, observation optical fundus is unaffected, II grade: vitreous body moderate is muddy, still can see optical fundus, III grade through muddiness: vitreous body severe is muddy, optical fundus be can't see, the incidence rate of I, II, III grade added up, experimental result show sample group vitreous opacity mild degree compared with matched group, has significant difference.

embodiment 17 pharmaceutical carrier beneficial effect (minimizing drug toxicity)

(1) get healthy new zealand white rabbit 10, male and female are not limit, and body weight 1.5 ~ 2.0kg, is divided into A at random, B two groups, often organize 5;

(2) the left ear auricular vein of A group rabbit slowly injects the pharmaceutical carrier 10mL having wrapped up paclitaxel obtained in embodiment 14, and the left ear auricular vein of B group rabbit slowly injects the paclitaxel injection of same concentrations in contrast, all injects once every day for two groups;

(3) rabbit was put to death after one week by injection continuously, clip injection site ear, put into 10% formalin and fixed, paraffin embedding, section, HE dyeing, the change of light Microscopic observation blood vessel and the downright bad situation of tissue;

(4) result shows, with biomembrane parcel Taxol injection after, rabbit auricular vein injection site blood vessel structure is complete, vascular endothelial cell without swelling, degeneration, necrosis, tube wall NIP cellular infiltration; And with after common paclitaxel injection injection, rabbit auricular vein injection site blood vessel structure is damaged, vascular endothelial cell swelling, partial denaturation and necrosis, and tube wall has inflammatory cell infiltration.

embodiment 18 pharmaceutical carrier beneficial effect (improving long-lasting)

(2) calcitonin is dissolved in PBS, adds the biomembrane in ether dissolution step (1), and mix with above-mentioned calcitonin equal-volume, be placed in 50 DEG C of water-baths and hatch 20min, be and required wrapped up the pharmaceutical carrier of calcitonin;

(3) the methanol ultrasonic dissolution of the pharmaceutical carrier volume ratio 500 times of having wrapped up calcitonin by above-mentioned acquisition, 37 DEG C of nitrogen blow rear PBS and redissolve, and carry out assay by immunoluminescence method to calcitonin, and calculating calcitonin content in carrier is 8.6%;

(4) get healthy new zealand white rabbit 10, male and female are not limit, and body weight 1.5 ~ 2.0kg, is divided into A at random, B two groups, often organize 5;

(5) the pharmaceutical carrier 10ml having wrapped up calcitonin of the above-mentioned acquisition of A group intravenous injection, the calcitonin of B group injection same medicine dosage is in contrast;

(6) result shows, with the calcitonin of biomembrane parcel in vivo drug half-life be 4 hours, and the common calcitonin half-life is 2 hours, is long-lastingly doubled.

embodiment 19 pharmaceutical carrier beneficial effect (medicine passive targeting/reduction toxicity)

(1) under 4 DEG C of conditions, amphotericin B is dissolved in ethanol, form saturated solution;

(2) the cellular compartment mixing will obtained in above-mentioned amphotericin B saturated solution and embodiment 5, use sonicator ultrasonic emulsification in ice bath environment, the amplitude of ultrasonic probe is 45%, and work 30sec, interval 30sec, circulate 20 times;

(3) by emulsion 100,000 × g, 20min, centrifugal under 4 DEG C of conditions, abandon supernatant, collecting precipitation, be and required wrapped up the pharmaceutical carrier of amphotericin B;

(4) the pharmaceutical carrier 5%Triton X-100 having wrapped up amphotericin B by above-mentioned acquisition jiggles at 37 DEG C the breakdown of emulsion that spends the night, and with the methanol dilution of volume ratio 100 times, carry out assay by high performance liquid chromatography to amphotericin B, the content calculating amphotericin B in carrier is 8.3%;

(5) get healthy mice 30, male and female are not limit, body weight 18-22g, are divided into A at random, B two groups, often organize 15;

(6) the above-mentioned pharmaceutical carrier 0.2mg having wrapped up amphotericin B of A group mouse tail vein injection, the amphotericin B of B group injected in mice same medicine dosage in contrast, respectively at after administration 0.5,1.0,1.5,2.0 and each group of 5.0h get the blood sampling of 3 mouse hearts after put to death, and take out the internal organs such as liver,kidney,spleen and carry out amphotericin B assay;

(7) result shows, is obviously less than common amphotericin B, effectively reduces nephrotoxicity by the drug distribution of amphotericin B in kidney of biomembrane parcel.

show amphotericin B concentration (ug/g) in different time mice serum and internal organs

embodiment 20 pharmaceutical carrier beneficial effect (drug main moving-target tropism)

(1) Human Hepatic Carcinoma Cell Line BEL-7402 is inoculated in BALB/C nude mice dorsal sc, when diameter of tumor is about 1cm, get the fresh tumor tissue of flesh of fish sample and be cut into 1mm*2mm tumor block, implant in the tunnel under the left outside leaf peplos of nude mice with ophthalmic tweezers, cotton swab is sewed up after gently pressing hemostasis and is closed abdomen;

(2) 12d exploratory laparotomy after modeling, 20 model mouses selecting tumorous size basically identical are divided into A at random, B two groups, often organize 10, the amycin pharmaceutical carrier intraperitoneal injection every day 0.2ml with targeting that A group will obtain in embodiment 12, B group by the amycin that obtains on the market in contrast every day intraperitoneal injection 0.2ml;

(3) after 10d, model mouse is opened abdomen and measure tumor size, and tumor tissues be used for pathological section and put through observing, the tumor control rate of result show sample group is higher than matched group.

embodiment 21 transgenic beneficial effect (transgenic effectiveness)

(1) Chinese hamster ovary celI is inoculated in 6 orifice plates, with DMEM complete medium (containing serum) at 37 DEG C, 5% CO ₂24h is cultivated under condition;

(2) transgene carrier having wrapped up hepatitis B virus DNA plasmid obtained in embodiment 7 is joined in the DMEM culture medium of serum-free, and mix homogeneously gently;

(3) the above-mentioned serum-free DMEM culture fluid containing hepatitis B virus DNA transgene carrier is applied on Chinese hamster ovary celI uniformly, and in cell culture incubator (37 DEG C, 5% CO ₂) cultivate the DMEM culture fluid added again after 6h containing 20% hyclone, after continuing to cultivate 48h, change liquid with the DMEM culture fluid containing 10% hyclone;

(4) after cell transfecting 48h, in culture fluid, hbs antigen can be detected, transgenic success.

embodiment 22 transgenic beneficial effect (raising transgene efficiency)

(1) plasmid containing hepatitis B virus DNA is dissolved in the PBS of 20mmol/L, and mixes with commercially available lipofectamine;

(2) Chinese hamster ovary celI containing the transfection of hepatitis B virus DNA transgene carrier will be used in embodiment 21 as A group, B group is the Chinese hamster ovary celI with the liposome transfection of the commensurability hepatitis B virus DNA of A group that contains obtained in above-mentioned steps (1);

(3), after cell transfecting 48h, the content of hbs antigen in culture fluid is detected;

(4) result as Fig. 1 shows, with biomembrane as genetically modified transfection reagent, effectively can improve transgene efficiency.

embodiment 23 transgenic beneficial effect (minimizing cytotoxicity)

(1) trypsinization is used by after the cell transfecting 72h of A group and B group in embodiment 22, through Flow cytometry different transfection group apoptosis situation after PI dyeing;

(2) result shows, with biomembrane as genetically modified transfection reagent, apoptosis rate is significantly less than liposome as transfection reagent, effectively can reduce cytotoxicity.

embodiment 24 cosmetics additive beneficial effect (moisturizing)

(1) random choose ten people, everyone selects the position of 10cm × 10cm in left and right the back of the hand center, and any hands smears the biomembrane of redissolution in 500ul volume PBS of acquisition in embodiment 3, while the PBS smearing same volume is in contrast;

After (2) two palmistry co-located smear two weeks continuously, by the water content of skin moisture tester test both sides skin;

(3) result display, smearing biomembranous the back of the hand mean skin water content is 48%, and the back of the hand mean skin water content of smearing PBS is 43%, and biomembrane effectively can carry out skin moisture-keeping as cosmetics additive.

embodiment 25 cosmetics additive beneficial effect (whitening)

(1) with 0.5 mmol/L L-DOPA for substrate, in the survey live body system of 1mL 0.05mol/L phosphate buffer, first add the biological compartment (being dissolved in DMSO solution) of acquisition in 20uL embodiment 4 in the pear-shaped tube of 1.5mL, add 400uL more in advance at the substrate solution of 30 DEG C of water bath with thermostatic control insulations, with buffer solution, volume is added to 970uL, then 30uL tryrosinase aqueous solution is added, mix at once, take PBS as contrast, under 30 DEG C of constant temperature, measure the light absorption value growth straight line in time when wavelength is 475nm in 1min, try to achieve enzyme activity from straight slope;

(2) result shows, biological compartment is 37% to the inhibit activities of tryrosinase, and the inhibit activities of the tryrosinase of matched group is obvious hardly, and biomembrane effectively can promote whitening as cosmetics additive.

embodiment 26 cosmetics additive beneficial effect (defying age)

(1) random choose 45 years old to 55 years old age bracket ten people, everyone selects the position of 10cm × 10cm in left and right the back of the hand center, the hands on any one side smears the biomembrane of redissolution in 500ul volume PBS obtained in embodiment 3, smears the PBS of same volume in contrast;

After (2) two palmistry co-located smear surrounding continuously, by the defying age situation of skin elasticity tester test both sides skin;

(3) result display, smearing biomembranous the back of the hand mean skin elasticity is 30%, and the back of the hand mean skin elasticity of smearing PBS is 27%, and biomembrane can effective defying age as cosmetics additive.

embodiment 27 cosmetics carrier beneficial effect (improving the stability of functional components)

With embodiment 13.

embodiment 28 cosmetics carrier beneficial effect (improving the dissolubility of functional components)

(1) under 4 DEG C of conditions, Rhizoma Curcumae Longae extract is dissolved in butanediol, forms saturated solution;

(2) the cellular compartment mixing will obtained in above-mentioned curcumin saturated solution and embodiment 5, use sonicator ultrasonic emulsification in ice bath environment, the amplitude of ultrasonic probe is 40%, and work 30sec, interval 30sec, circulate 20 times;

(3) by emulsion 100,000 × g, 20min, centrifugal under 4 DEG C of conditions, abandon supernatant, collecting precipitation, be and required wrapped up the pharmaceutical carrier of curcumin;

(4) the pharmaceutical carrier PBS having wrapped up curcumin by above-mentioned acquisition is resuspended to 1ml, and jiggles at 37 DEG C the breakdown of emulsion that spends the night with 5%Triton X-100, and with the methanol dilution of volume ratio 100 times;

(5) carry out assay by high performance liquid chromatography to curcumin, calculating in 1ml solution curcumin content in carrier is 10.9%;

(6) excessive curcumin is put in PBS buffer solution, jiggle for 37 DEG C, the changes of contents of sampling and measuring curcumin, no longer changes to concentration at set intervals, adopt high performance liquid chromatography, calculate the content of curcumin for 4.0ug/mL with external standard counter point;

(7) result shows, the dissolubility of the curcumin wrapped up by biomembrane improves more than 2000 times.

embodiment 29 cosmetics carrier beneficial effect (improving the long-lasting of functional components)

(2) Symwhite-337 is dissolved in ethanol, add the biomembrane in ether dissolution step (1), and mix with the volume ratio of 1:10 with above-mentioned Symwhite-337, be placed in 50 DEG C of water-baths and hatch 20min, be and required wrapped up the cosmetics carrier of Symwhite-337;

(3) by above-mentioned acquisition wrapped up the cosmetics carrier breakdown of emulsion of Symwhite-337 after by high performance liquid chromatography assay is carried out to Symwhite-337, being diluted to concentration with PBS is 0.5%;

(4) choose the skin of a place 10cm × 10cm, smear the cosmetics carrier 1ml having wrapped up Symwhite-337 that above-mentioned steps (3) obtains, every day smears, sooner or later respectively once;

(5) as shown in Figure 2, result shows, can long-acting whitening with the Symwhite-337 of biomembrane parcel.

embodiment 30 cosmetics carrier beneficial effect (improving the skin permeation rate of functional components)

(1) by rat with after 2.5% Nembutal sodium solution intraperitoneal anesthesia, back hair is carefully shaved off with shaver, take off plucked skin, reject subcutaneous fat and muscle, the dermatological specimens of diameter 22mm made by the rat skin in vitro card punch prepared, and be placed in Frnaz diffusion cell, epidermis side is to supply pool, acceptance pool volume is 14.5mL, effective infiltrating area 0.785cm ², acceptable solution is 0.9%Nacl solution;

(2) join in supply pool by biomembrane by electrostatic surface absorption method absorption collagen protein, the collagen protein of same amount in contrast;

(3) keep 37 DEG C of constant temperature in experimentation, have magnetic stirrer in reception tank, mixing speed 200r/min, respectively 1,2,3,5,7,9h draws 200ul sample from reception tank, supplements the fresh acceptable solution of same volume immediately simultaneously;

(4) measure sample fluorescence value with fluorophotometer, substitute into the transdermal concentration of regression equation calculation medicine Different periods;

(5) as shown in Figure 3, result shows, wraps up collagen protein with biomembrane as carrier, can significantly improve its transdermal absorption factor.

embodiment 31 cosmetics carrier beneficial effect (improving the effectiveness of functional components)

(1) B16 melanocyte is inoculated in 6 orifice plates, with DMEM complete medium (containing serum) at 37 DEG C, 5% CO ₂cultivate 24h under condition, abandon supernatant;

(2) the ascorbic carrier that wrapped up obtained in embodiment 8 is joined in the DMEM culture medium of serum-free, and mix homogeneously gently, usual vitamin C is in contrast;

(3) the above-mentioned serum-free DMEM culture fluid containing vitamin C carrier is applied on B16 melanocyte uniformly, and in cell culture incubator (37 DEG C, 5% CO ₂) cultivate after 3d, abandon supernatant, PBS washs, and every hole adds pancreas peptic cell 5min, respectively to often organizing cell counting;

(4) cell suspension is 20,000 × g, 15min, centrifugal under 4 DEG C of conditions, abandons supernatant, and precipitation adds 1mol/L NaOH solution, and 80 DEG C of heating 30min, detect the absorbance under 475nm with spectrophotometer;

(5) pass through formula: B16 cell suppression ratio=[1-(medicine hole absorbance/medicine porocyte number)/(control wells absorbance/control wells cell number)] × 100%, calculate obtain in embodiment 8 to have wrapped up ascorbic carrier B16 cell suppression ratio be 38.5%.(Fig. 4)

embodiment 32 vaccine carrier and adjuvant beneficial effect (safety)

For determining whether biomembrane exists the material that can cause allergic reaction as the carrier of vaccine or immunological adjuvant, guarantees the safety of biomembrane vaccination, invention has been animal hypersensitive test.

(1) by embodiment 1, embodiment 2, embodiment 3, embodiment 4, the biomembrane obtained in embodiment 5 inoculates Cavia porcellus respectively, often organizes 2, every Cavia porcellus subcutaneous vaccination 1mL, one week, interval, subcutaneous vaccination second time, makes Cavia porcellus sensitization, with same sample every intravenous injection 500ul after three weeks;

(2) result judges: after intravenous injection, and the phenomenon of dysphoria, shock or death does not all appear in Cavia porcellus, illustrates that injection mass there is no anaphylaxis to Cavia porcellus.

the application (prevention viral vaccine, subcutaneous dosage form) of embodiment 33 vaccine

(1) choose a collection of 42d, ternary healthy, of uniform size is mixed little pig's head, is divided into A at random, B two groups, often organizes 20;

(2) the vector immunity A group of having wrapped up swine fever virus that will obtain in embodiment 9, in contrast, two groups of test pig are raised under identical feeding and management condition to use conventional swine Fever Vaccine immunity B group on the market, by cephalic vein blood sampling 3ml after 42d, separation of serum;

(3) serum carries out the detection (as following table) of hog cholera antibody ELISA, and immunization rate is 95%.

the application (gene vaccine, intramuscular injection dosage form) of embodiment 34 vaccine

(1) beasle dog 10 is chosen, average weight 3kg(2.5 ~ 3.5kg), male and female are not limit, and are divided into A at random, B two groups, often organize 3;

(2) the biomembrane intramuscular injection A group of canine distemper virus DNA plasmid will have been wrapped up, every only injection 500ul plasmid DNA concentration is the biomembrane vaccine of 1ug/u1, the canine distemper virus DNA plasmid of B group injection same DNA concentration in contrast, injected a vaccine every two weeks;

(3) diluent of canine distemper morbidity dog brain tissue homogenate 1:10 process with dual anti-after immune two weeks of third time, carry out 2ml challenge viral dosage often to all experimental dogs, wherein 1ml collunarium, 1ml is intramuscular injection;

(4) after counteracting toxic substances, every day surveys its rectal temperature, the morbidity of itemized record experimental dog and death condition, observes the clinical symptoms change of animal;

(5) after counteracting toxic substances there is death condition in B group 2 experimental dogs, and remaining 1 occurs spared nerve symptom, and most high fever reaches 41 DEG C, then slowly lapses to normal.Also there is rising in various degree in biomembrane injection group body temperature, is up to 39.4 DEG C, but does not occur nervous symptoms and death;

(6) result shows, biomembrane has very strong immune effect as the carrier of gene vaccine and immunological adjuvant.

the application (tumor vaccine, peroral dosage form) of embodiment 35 vaccine

(1) get healthy mice 10, male and female are not limit, body weight 18-22g, are divided into A at random, B two groups, often organize 5;

(2) before immunity and inoculation, mice fasting water more than 12 hours, 30min first per os feed 150ul 0.01mol/L NaHCO before inoculation ₃solution with in and gastric acid, the biofilm carrier vaccine per os feed A group of Helicobacter pylori recombiant protein will have been wrapped up, every 100ul Helicobacter pylori recombiant protein concentration of only feeding is the biomembrane vaccine of 10ul, and B group is fed 100ul PBS, wherein containing Helicobacter pylori recombiant protein 10ul in contrast;

(3) experimental group and matched group are each immunity in the 0th, 7,14,21 day once, and immunity recovered drinking water after 1 hour, (the 35th day) each group all use 10 apart from final immunization after 2 weeks ⁸the Helicobacter pylori of CFU/ time is attacked, the next day once, totally three times, after last attacks 4 weeks, put to death mice;

(4) cut open the belly immediately after putting to death mice and get stomach, get spleen, cut off along large curved side, rinse gastric content with physiological saline solution, get gastric tissue along the longitudinal axis, the capable rapid urease test of rearmounted 96 orifice plate of smear;

(5) urease test display; biomembrane vaccine group and matched group compare; effectively can not be subject to the infection of Helicobacter pylori by Protection Mus; infection due to Helicobacter pylori is the important paathogenic factor of the disease such as gastric cancer, mucosa tissue lymphoma; therefore, biomembrane is used effectively can to prevent the diseases such as gastric cancer as the vaccine carrier of Helicobacter pylori recombiant protein and adjuvant.

embodiment 36 vaccine carrier and adjuvant beneficial effect (potent humoral immunity)

With embodiment 33

embodiment 37 vaccine carrier and adjuvant beneficial effect (strong cellular immunity)

(1) expression mouse spleen obtained in embodiment 35 being carried out splenocyte INF-γ and IL-4 mRNA compares;

(2) result display, use biomembrane as the vaccine carrier of Helicobacter pylori recombiant protein and adjuvant, mouse spleen lymphocyte occurs based on the hypertrophy of Th1 cell (INF-γ), and INF-γ level comparatively matched group compares and has significant difference.

the application of embodiment 38 artificial organ

(1) 2 monthly age beasle dog 10 is chosen, average weight 2.5kg(2 ~ 3kg), male and female are not limit, and are divided into A at random, B two groups, often organize 5;

(2) by right for beasle dog knee joint from outer lateral incision capsula articularis genus, do not cut off side para-ligament, lateral meniscus done a triangle defect (base periphery, most advanced and sophisticated inwardly to the half of meniscus body width), then makes a longitudinal deficiency in ischemic region;

(3) the artificial meniscus that A group will obtain in embodiment 10 implants defect, and B group is that the artificial meniscus of support implants defect in contrast with collagen by what obtain on the market, all sews up fixing with 5-0 absorbable thread for two groups, after sew up the incision, postoperatively to raise in cages;

(4) postoperative in 12 weeks execution animals, get knee joint specimen to observe: the visible defect place healing of A group is good, the new tissue growth of defective region adularescent, its quality, color and luster its nothing similar to the meniscal tissue of surrounding normal is significantly demarcated, B group visible implant edge quality, color and luster are close with the normal meniscus, defective region visible white fibrous scar sample tissue, corresponding femur and tibial prosthesis face show slightly coarse.

Claims

1. the biomembrane, the closing structure with biomembrane or the cellular compartment that obtain of natural origin and/or self-assembling technique, it is characterized in that: the biogenetic derivation of this biomembrane, the closing structure with biomembrane or cellular compartment is animal, plant or microorganism, the biomembrane obtained is complete or fragment, for being lipid bilayer Rotating fields in form, its constituent is lipid and protein mainly, separately has a small amount of saccharide by being covalently bonded on lipid or protein.

2. the biomembrane, the closing structure with biomembrane or the cellular compartment that obtain of natural origin and/or self-assembling technique according to claim 1, it is characterized in that: described biomembrane, the closing structure with biomembrane or cellular compartment, particle diameter is from 10nm to tens microns.

3. the biomembrane, the closing structure with biomembrane or the cellular compartment that obtain of natural origin and/or self-assembling technique according to claim 1, is characterized in that: described biomembrane, the shape of closing structure or cellular compartment with biomembrane comprise the variform structure of spherical, vesicle shape, shaft-like, spiral helicine monolayer or multilamellar, multicell.

4. the biomembrane, the closing structure with biomembrane or the cellular compartment that obtain of natural origin and/or self-assembling technique according to claim 1, is characterized in that: described biomembrane comprise plasma membrane, nuclear membrane, mitochondrial membrane, endoplasmic reticulum, lysosome membrane, Golgi membrane, chloroplast membranes and vacuole and peroxisome film one or more.

5. the biomembrane, the closing structure with biomembrane or the cellular compartment that obtain of natural origin and/or self-assembling technique according to claim 1, is characterized in that: described cellular compartment refers to general organelle; As preferably, described cellular compartment be mitochondrion, chloroplast, peroxisome, lysosome, endoplasmic reticulum, nucleus, Golgi body and vesicle and microtubule one or more.

6. the biomembrane described in claim 1 ~ 5 any one claim, there is the closing structure of biomembrane or a preparation method for cellular compartment, it is characterized in that the method comprises the following steps:

1) biological cell is obtained;

7. preparation method according to claim 6, is characterized in that differential centrifugation extracting method comprises the following steps:

8. preparation method according to claim 6, is characterized in that density gradient centrifugation extracting method comprises the following steps:

9. the preparation method stated according to Claim 8, is characterized in that the mass percent concentration scope of step 1. sucrose solution is 10 ~ 70%; As preferably, the different quality percent concentration of described step 1. sucrose solution is 10%, 20%, 30%, 40%, 45%, 50%, 55%, 60%, 65%, 70%.

10. preparation method according to claim 6, is characterized in that two-phase extracting process comprises following feature:

11. preparation methoies stated according to claim 10, is characterized in that described two-phase is the biphase mixture of glucosan/Polyethylene Glycol.

12. preparation methoies stated according to claim 10, it is characterized in that described two-phase comprise water two-phase or, organic biphasic, aqueous phase solution and organic phase solution, described solvent is selected from any one or its combination of water, acetonitrile, acetone, oxolane, methanol, ethanol, propanol.

13. 1 kinds of self assemblies are prepared biomembrane, are had the method for the closing structure of biomembrane or cellular compartment, it is characterized in that the method comprises the preparation method of claim 6 ~ 12, and biomembrane step 3) obtained, there is the closing structure of biomembrane and the material of cellular compartment covers on wall of a container with the form of desciccator diaphragm, then slowly water or buffer solution is injected, and slight or violent vibration, biomembrane, the closing structure with biomembrane and cellular compartment needed for self assembly obtains.

14. preparation methoies according to claim 13, is characterized in that step 3) obtained material dissolves in chloroform solvent, join in container, reduction vaporization makes biomembrane sprawl on vessel surface, add PBS buffer solution after being evaporated to constant weight and slowly shake 0.5 ~ 3h, in 100,000 ~ 200,000 × g, 30 ~ 90min, ultracentrifugation under 1 ~ 6 DEG C of condition, abandons supernatant, collecting precipitation, is required biomembrane, has the closing structure of biomembrane and cellular compartment.

Biomembrane, the closing structure with biomembrane or cellular compartment described in 15. claim 1 ~ 5 any one claim application, this is applied as described biomembrane, have the closing structure of biomembrane or cellular compartment to wrapping up in the film of active component, in film parcel, surface adsorption, surface-crosslinked, intermembranous inlay and wrap up in film add targeting.

16. application according to claim 15, is characterized in that: active component comprises vaccine or immunomodulatory activity composition, cosmetics or cosmetic active ingredient, active constituents of medicine, genetic stew and cell or tissue.