CN106279439A - A kind of oligopeptide 1 fusion protein containing cell-penetrating peptide and preparation method thereof - Google Patents
A kind of oligopeptide 1 fusion protein containing cell-penetrating peptide and preparation method thereof Download PDFInfo
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- CN106279439A CN106279439A CN201610728673.8A CN201610728673A CN106279439A CN 106279439 A CN106279439 A CN 106279439A CN 201610728673 A CN201610728673 A CN 201610728673A CN 106279439 A CN106279439 A CN 106279439A
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- fusion protein
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- 102000020313 Cell-Penetrating Peptides Human genes 0.000 title claims abstract description 42
- 108010051109 Cell-Penetrating Peptides Proteins 0.000 title claims abstract description 42
- 108020001507 fusion proteins Proteins 0.000 title claims abstract description 25
- 102000037865 fusion proteins Human genes 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 108010038807 Oligopeptides Proteins 0.000 title abstract description 5
- 102000015636 Oligopeptides Human genes 0.000 title abstract description 5
- 239000000203 mixture Substances 0.000 claims abstract description 23
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- 241000588724 Escherichia coli Species 0.000 claims abstract description 6
- 150000001875 compounds Chemical class 0.000 claims abstract description 3
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 16
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 14
- 239000000872 buffer Substances 0.000 claims description 12
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- 239000006166 lysate Substances 0.000 claims description 6
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/485—Epidermal growth factor [EGF], i.e. urogastrone
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Gastroenterology & Hepatology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses industrialized production and contain ELP (Val Pro Gly Xaa Gly) cell-penetrating peptide and the fusion protein of oligopeptide 1 (EGF), compositions containing this fusion protein, and the preparation method of this fusion protein, use escherichia coli by seed culture, expand fermentation, thalline collection, broken, cracking, the process such as isolated and purified collect oligopeptide 1 fusion protein containing cell-penetrating peptide.The present invention has the advantages such as method is simple, production concentration is high, running cost is low, is suitable for industrialized production.The compound of the present invention and compositions may be used as active component or the additive of medical treatment consumptive materials, health product and cosmetics.
Description
Invention field
The present invention relates to oligopeptide-1 that a kind of industrialized production contains cell-penetrating peptide, i.e. the preparation method of PHPV gene protein,
Belong to bioengineering field.
Background of invention
Cell-penetrating peptide (CPP) is the little molecule multiple stage having and penetrating various kinds of cell film function, can carry biological activity big
Molecular substance enters cell.Some found in recent years have the small peptide of cell-penetrating function, less than 30 aminoacid, i.e. cells
Cell-penetrating peptide CPPs, it is possible to effectively protein, polypeptide, nucleic acid fragment etc. are imported mammalian cell in many ways, its tool
There is transduction efficiency high and do not result in the feature of cell injury.Up to the present, it has been found that multiple cell-penetrating peptide, they are common
Have a characteristic that 1. there is clean positive charge and amphipathic；2. film transport efficacy is worn high；3. can import nearly all thin
Born of the same parents；4. can carry various active material and enter cell；5. can targeting permeates cell membranes, reach inactivate multiple virus and do not cause
Cell injury.According to the literature, WENDER P A, waits (WENDE P A, ROTHBARD J B, JESSOP T C, et
al.Oligocarbamate molecular transporters:Design,synthesis,and biological
evaluation of a new class of transporters for drug delivery.JAme Chem Soc,
2002,124 (45): 13382-13383) one is devised containing cell-penetrating peptide, elastin laminin similar polypeptide ELP (Val-Pro-
Gly-Xaa-Gly) and a kind of Binding peptide being derived from cyclin-dependent kinase inhibitors P21, this peptide species can
The growth rate making SKOV-3 ovarian cancer cell and Hela cervical cancer cell slows down and suppresses their propagation.
Connection peptides is to connect each functional domain component in protein so that form each functional structure of this protein molecule
Territory keeps its activity conformation, plays its respective biological function, and the performance for each functional domain biologic activity provides association
Together, regulation and allostery, it is to avoid because the factors such as electric charge/space potential barrier cause the bioactive forfeiture of protein molecule.Generally
Connection peptides is divided into elasticity, rigidity, three kinds of forms of cuttability.Connection peptides typically has multiple secondary structure state to play it
Biological function, such as α spiral, β-pleated sheet chain, crimps, bends and corner.
Oligopeptide-1 has another name called epithelical cell growth factor) it is a kind of active substance in human body, the work being made up of 53 amino
Property polypeptide.Oligopeptide-1 be United States Biochemical scholar Stanley doctor Cohen in 1962 at first at the extracting solution of mice lower jaw gland
Middle discovery, and propose this factor and have a growth directly facilitating Mucocutaneous reparation and epidermis cell, and the cell of aging can be made
Against being divided into young cell, there are promotion collagen protein and the synthesis of elastin, can promote that granulation tissue regenerates, delay cell to decline
Always, nourish female sex organ, the effect of desalination wrinkle and mottle, it be physianthropy and greatest discovery in History of Aesthetics it
One.The EGF somatomedin that Stanley doctor Cohen finds is in 1986 Nian Huo world Nobel Prize in Physiology or Medicines.Oligopeptide-1 mat
By the tyrosine phosphorylation of the few peptide-1 receptor of stimulation, reach to repair hypertrophy skin surface cell, the table impaired to wound, ulcer etc.
Musculus cutaneus skin has excellent curative effect, tract mucosa is had moisturizing, compact, the effect of self-cleaning.Its maximum feature is can to promote carefully
The inverse differentiation of the propagation of born of the same parents, thus replace old and feeble and dead cell with newborn cell.Initial oligopeptide-1 is mainly employed for
Medical domain, to promote reparation and the regeneration of impaired epidermis, such as treatment burn, scald, ulcer, promotion wound healing, repairs intestinal
The damage etc. of gastropore, liver and cornea, effect is the most notable.Oligopeptide-1 expansion is applied to medical treatment consumption by modern clinic already
The cosmetic field of material, body-care and defying age.Thus, it is big that the preparation synthesis of oligopeptide-1 faces world market demand, produces
The problem that product resource is the most rare, in order to solve the world market demand supplies to oligopeptide-1EGF dressing, my associating U.S. of company
Harvard University doctor Wei, Shandong Agricultural University doctor Yu, China biological medicine academy, Guangdong medical insurance pharmaceutcal corporation, Ltd are common
Research, initially with disclosed cell-penetrating peptide ELP structures such as WENDER P A, constructs a kind of with the yeast expressed widow containing ELP
Peptide-1EGF preparation method, its prepared high-purity obtained merges new albumen, i.e. PHPV gene protein, not only has cell-penetrating peptide
ELP and the multiple-effect function of oligopeptide, and negative charge (anion) the C end of the new protein surface after its crosslinking fusion and human papilloma
N on virus HPV granule rectifies electric charge (cation) mutually complexation, and the N end positively charged by its surface is dredged with middle
The C end negative charge in pool and the site, cog region containing peptidase combines, and film indexing signal is even with the NLS of nuclear localization signal
Connection, thus blocks human papillomavirus and invades in host cell, then by the interaction of cell membrane permeates cell membranes this
The natural cover for defense, plays break virus after birth shell and carrys out inactivation of bacterial virus, reaches prevent precancerous lesions of uterine cervix and realize treating HPV
The purpose infected.The oligopeptide-1 containing cell-penetrating peptide is the most not yet had to merge new albumen genetic engineering work in escherichia coli at present
Industry production method.
Summary of the invention
Present invention aim to address that industrial gene engineering colibacillus produces melting of the oligopeptide-1 (EGF) containing cell-penetrating peptide
The method closing new albumen.
In order to achieve the above object, the invention provides following inventive concept.
Present invention firstly provides a kind of oligopeptide-1 including cell-penetrating peptide ELP and merge new albumen, i.e. PHPV gene protein.
Wherein, cell-penetrating peptide ELP is positioned at the C-end of oligopeptide-1；
Preferably, cell-penetrating peptide ELP is connected with the C-end of oligopeptide-1 by connection peptides；
Wherein, connection peptides preferably crimps connection peptides, and including (GGGGS) N, wherein N is greater than or equal to the integer of 1；
Preferably, the C-end of cell-penetrating peptide ELP next-door neighbour oligopeptide-1；
Wherein, the sequence of cell-penetrating peptide ELP is: Val-Pro-Gly-Xaa-Gly；
Preferably, Xaa is one of S, E, V, A, V, preferably A.
Present invention also offers a kind of targeting selectivity administration composition, said composition includes cell-penetrating peptide ELP and oligopeptide-1
Fusion protein；
Optional, said composition also includes excipient, stabilizer or diluent.
The invention still further relates to the preparation method of the fusion protein of the oligopeptide-1 that a kind of industrialized production contains cell-penetrating peptide ELP,
It comprises the steps:
(1) prepared by seed liquor；
(2) Escherichia coli fermentation；
(3) microorganism collection crushes；
(4) oligopeptide-1 fusion protein purification containing cell-penetrating peptide ELP；
(5) compounding targeting selectivity administration composition.
Wherein, the lysate used in described microorganism collection destruction step is 20mM trishydroxymethylaminomethane, 8M carbamide
The buffer formed；
Wherein, the purification of described fusion protein is to use the exchange of buffer solution, ion, the compound mode of gel chromatography.
Cell-penetrating peptide ELP employing WENDE P A used in the present invention etc. (WENDE P A, ROTHBARD J B,
JESSOP T C,et al.Oligocarbamate molecular trans porters:Design,synthesis,and
biological evaluation of a new cla ss of transporters for drug delivery.JAme
Chem Soc, 2002,124 (45): 13382-13383) method design disclosed in obtains, and combines the spy of oligopeptide-1 polypeptide fragment
Point, has carried out checking selection to the kind of ELP the 4th amino acids residue, has found when Xaa is S, E, V, A, V, or the fusion obtained
Albumen has stronger targeting, and particularly when Xaa is A, the usefulness of its fusion protein obtained is optimum, and this fusion protein contains
There is the fragments of peptides of more positive charge, utilize the principle of positive and negative charge non-specific interaction to be combined with cell membrane, open skin
Skin cell membrane thus by destination protein targeting import skin base layer receptor.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are closely for illustrating the present invention
Rather than restriction the scope of the present invention.The experimental technique of unreceipted actual conditions in the following example, generally according to conventional strip
Part, or according to the condition proposed by manufacturer.Ratio and percentage ratio are based on weight, unless stated otherwise.
Embodiment 1: prepared by the fusion protein of the oligopeptide-1 (EGF) containing cell-penetrating peptide ELP
(1) prepared by seed liquor:
During picking list bacterium colony accesses 5-15ml seed culture medium from-70 DEG C of strain tubes preserved, it is simultaneously introduced ammonia benzyl blue or green
Mycin so that it is concentration in the medium reaches 50-200mg/ml, 22 32 DEG C, it is little that 150 250rpm shaking tables cultivate 8 12
Time, by 1 10% inoculum concentration, culture fluid above is accessed in seed culture medium again, is simultaneously introduced ampicillin so that it is
At 27 32 DEG C, 150 250rpm shaking tables cultivate 8 12 hours.
(2) Escherichia coli fermentation
Aseptically, above-mentioned cultured seed liquor is inoculated in fermentation medium by 30ml inoculum concentration again；Send out
Ferment medium component (g/L): tryptone 10g, yeast powder 5g, sodium chloride 10g, ampicillin 0.1g deionized water dissolving
Constant volume, autoclaving；Fermentation condition: temperature 30-32 DEG C, fermentation to OD600 reaches the different of about 0.8 addition derivant 100mM
Propyl dithiocarbamate galactoside (IPTG) 0.1% is induced, and collects thalline after 34 hours.
(3) microorganism collection crushes:
By fermentation liquid with speed centrifuge 20 30min of 6000 12000rpm, abandon supernatant and retain precipitation.
Taking precipitation, rinse 12 times with phosphate buffer (pH8.0), with 5 times of Tris-Hcl buffer solution, 1800w supersound process is broken
Broken, then with refrigerated centrifuger, buffer is abandoned supernatant with the centrifugation of 6000 12000rpm under the conditions of 2 18 DEG C, take
Precipitate and dissolve with 15 45 times of lysates, the buffering that lysate used is 20mM trishydroxymethylaminomethane, 8M carbamide is formed
Liquid, under the conditions of 2 18 DEG C, centrifugal 30min collects supernatant.
(4) new protein purification is merged containing cell-penetrating peptide oligopeptide-1 (EGF)
Supernatant tomographic system is added in source 30S expansion bed with 1.5 8.5ml/min speed, uses
When 80mMTris buffer speed washing to wavelength is 280nm, uv absorption peak value is zero, then with containing 1M sodium chloride 80mMTris
Buffer, with 13% ratio eluting, collects the eluting peak merging new albumen containing cell-penetrating peptide oligopeptide-1 (EGF).
By tomographic system, eluent above 15 30ml is added to Superdex75 gel layer with the speed of 4 8ml/min
In analysis post, then by 15mM Arginine buffer with the speed eluting of 4 8ml/min until to go out peak complete, examine with ultraviolet detection
Surveying instrument to detect under wavelength 280nm, collect and retain absworption peak, collected volume is 300 500ml, will collect liquid in-20 DEG C
Freezen protective.
(5) complex composition
After freezing stock solution containing cell-penetrating peptide and oligopeptide-1 being redissolved, add excipient mannitol, stabilizer arginine, sea
Algae sugar, diluent deionized water, complex composition.Described compositions is medical apparatus and instruments compositions, Halth-care composition, cosmetics
Compositions.
Embodiment two: prepared by the fusion protein of the oligopeptide-1 (EGF) containing cell-penetrating peptide ELP
(1) escherichia coli spread cultivation fermentation:
During picking list bacterium colony accesses 10ml seed culture medium from-70 DEG C of strain tubes preserved, it is simultaneously introduced ammonia benzyl penicillium sp
Element so that it is concentration in the medium reaches 100mg/ml, 30 DEG C, 200rpm shaking table is cultivated 10 hours, by 5% inoculum concentration by front
The culture fluid in face accesses in seed culture medium again, is simultaneously introduced ampicillin so that it is at 100mg/ml, 32 DEG C, 200rpm
Shaking table is cultivated 10 hours, then joins in fermentation medium by 5% inoculum concentration by the seed liquor of activation, fermentation medium components
(g/L): tryptone 10g, yeast powder 5g, sodium chloride 10g, ampicillin 0.1g deionized water dissolving constant volume, high pressure goes out
Bacterium；35 DEG C, 200rpm, use Tris-Hcl buffer to make pH value maintain 9.0, cultivate 10 hours, then fermentation temperature is improved
To 40 DEG C of inductions, continue to cultivate 4 hours.The oligopeptide-1 (EGF) of cell-penetrating peptide is expressed in strain.
(2) microorganism collection crushes:
With refrigerated centrifuger by under the conditions of fermentation liquid and 4 DEG C with the centrifugation 30min of 10000rpm, abandon supernatant retain
Save backup under the conditions of being deposited in 4 DEG C.Take acclimatization, by 10 times amount Tris-Hcl buffer solution, 900w supersound process 25
Individual circulation, each circulation 35min, then with refrigerated centrifuger by under the conditions of buffer and 4 DEG C with the centrifugation of 10000rpm
30min, abandons supernatant, takes precipitation and dissolves with 20 times of lysates, and lysate used is 20mM trihydroxy methyl amino first
The buffer that alkane, 8M carbamide are formed, the centrifugation 30min of 10000rpm, collects supernatant.
(3) new protein purification is merged containing cell-penetrating peptide oligopeptide-1 (EGF)
By tomographic system, this supernatant of 200ml is added in source 30S expansion bed with 3ml/min speed, use
80mMTris buffer washs to wavelength for uv absorption peak value during 280nm as zero with identical speed, then with containing 1M sodium chloride
80mMTris buffer, with 7% ratio eluting, collects the eluting peak merging new albumen containing cell-penetrating peptide oligopeptide-1 (EGF).
(4) gel separates:
By tomographic system, eluent above 25ml is added to Superdex75 gel chromatography column with the speed of 6.5ml/min
In, then by 15mM Arginine buffer with the speed eluting of 6.5ml/min until to go out peak complete, exist with ultraviolet detection detector
Detecting under wavelength 280nm, collect and retain absworption peak, collected volume is 350ml, will collect liquid in-20 DEG C of freezen protective.
(5) complex composition
After freezing stock solution containing cell-penetrating peptide and oligopeptide-1 being redissolved, add excipient mannitol, stabilizer arginine, sea
Algae sugar, diluent deionized water, complex composition.Described compositions is medical apparatus and instruments compositions, Halth-care composition, cosmetics
Compositions.
Embodiment 3: the film activity of wearing of oligopeptide-1 fusion protein containing cell-penetrating peptide ELP is verified
Utilize said method prepare the oligopeptide-1 containing cell-penetrating peptide i.e.: PHPV gene protein, innovative point is: (1) have send out
In ferment liquid, production concentration is high, more than 100mg/L, changes strain and expresses the recombination method of the oligopeptide containing cell-penetrating peptide, makes product table
Reach concentration and improve 1-2 times more in the past；(2) preparation technology is optimized: make technique extractive technique more simply (have only to 2 steps chromatographies
Operation, technique simplifies relatively beforeHalf)；(3) running cost is lower, purity is higher (more than 97%).The present invention be one more
Ripe industrialization innovation fermentation manufacturing technique, economic benefit is the most notable.
All above-mentioned primary these intellectual properties of enforcement, do not set this new product of enforcement limiting other forms
And/or new method.Those skilled in the art will utilize this important information, and foregoing is revised, to realize similar execution feelings
Condition.But, all modifications or transformation belong to the right of reservation based on new product of the present invention.
Claims (10)
1. oligopeptide-1 fusion protein including cell-penetrating peptide ELP, it is characterised in that described ELP is positioned at the C-end of oligopeptide-1.
Fusion protein the most according to claim 1, it is characterised in that described cell-penetrating peptide ELP is by connection peptides and oligopeptide-1
C-end is connected.
3. according to the fusion protein described in claim 1-2 any one, it is characterised in that described connection peptides is curling connection peptides,
Including (GGGGS) N, wherein N is greater than or equal to the integer of 1.
4. according to the fusion protein described in claim 1-3 any one, it is characterised in that described cell-penetrating peptide ELP next-door neighbour's oligopeptide-1
C-end.
5. according to the fusion protein described in claim 1-4 any one, it is characterised in that the sequence of described cell-penetrating peptide ELP is:
Val-Pro-Gly-Xaa-Gly。
6. according to the fusion protein described in claim 1-5 any one, it is characterised in that in described cell-penetrating peptide ELP Xaa be S,
One of E, V, A, V.
7. according to the fusion protein described in claim 1-6 any one, it is characterised in that in described cell-penetrating peptide ELP, Xaa is preferred
For A.
8. a targeting selectivity administration composition, said composition includes the fusion egg as described in claim 1-7 any one
In vain.
Compositions the most according to claim 8, it is characterised in that described compositions also includes excipient, stabilizer or dilution
Agent.
10. a preparation method for fusion protein as described in claim 1-7 any one, it comprises the steps:
(1) prepared by seed liquor；
(2) Escherichia coli fermentation；
(3) microorganism collection crushes；
(4) oligopeptide-1 fusion protein purification containing cell-penetrating peptide ELP；
(5) compounding targeting selectivity administration composition；
Preferably, in described preparation method, the lysate used in wherein said microorganism collection destruction step is 20mM tri-hydroxyl
The buffer that aminomethane, 8M carbamide are formed；
Or, in described preparation method, the purification of described fusion protein is to use the exchange of buffer solution, ion, gel layer
The compound mode of analysis.
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| CN106177911A (en) * | 2016-08-31 | 2016-12-07 | 易金阳 | A kind of biological composition dressing of prophylactic treatment HPV infection |
| CN107440986A (en) * | 2017-08-11 | 2017-12-08 | 烟台市华昕生物医药科技有限公司 | A kind of toothpaste |
| CN107827953A (en) * | 2017-11-08 | 2018-03-23 | 哈尔滨商业大学 | It is a kind of to target cell-penetrating peptide, the medicine with it and application |
| CN109385438A (en) * | 2018-11-23 | 2019-02-26 | 蓝脑科技(厦门)有限公司 | A kind of biofermentation preparation method of the active peptides with white-skinned face function |
| CN110448678A (en) * | 2018-05-07 | 2019-11-15 | 易金阳 | A kind of fusion factor containing PHPV is used to alleviate the component and Preparing Use of the eye drops of visual fatigue |
| CN110664742A (en) * | 2019-10-29 | 2020-01-10 | 易金阳 | Eye drops for relieving asthenopia and preparation method and application thereof |
| CN112625089A (en) * | 2020-12-14 | 2021-04-09 | 四川省恩乐生物工程有限公司 | Active protein peptide gene with excellent repairing effect and coded protein peptide |
| CN112955183A (en) * | 2018-09-25 | 2021-06-11 | 波尔多聚合技术研究所 | Bioconjugates of polysaccharides and elastin-like polypeptides and uses thereof |
| CN115154614A (en) * | 2022-07-08 | 2022-10-11 | 广西福莱明生物制药有限公司 | Fusion factor formula combination process |
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| CN106177911A (en) * | 2016-08-31 | 2016-12-07 | 易金阳 | A kind of biological composition dressing of prophylactic treatment HPV infection |
| CN107440986A (en) * | 2017-08-11 | 2017-12-08 | 烟台市华昕生物医药科技有限公司 | A kind of toothpaste |
| CN107827953A (en) * | 2017-11-08 | 2018-03-23 | 哈尔滨商业大学 | It is a kind of to target cell-penetrating peptide, the medicine with it and application |
| CN110448678A (en) * | 2018-05-07 | 2019-11-15 | 易金阳 | A kind of fusion factor containing PHPV is used to alleviate the component and Preparing Use of the eye drops of visual fatigue |
| CN112955183A (en) * | 2018-09-25 | 2021-06-11 | 波尔多聚合技术研究所 | Bioconjugates of polysaccharides and elastin-like polypeptides and uses thereof |
| CN112955183B (en) * | 2018-09-25 | 2024-04-30 | 波尔多聚合技术研究所 | Bioconjugates of polysaccharides and elastin-like polypeptides and uses thereof |
| CN109385438A (en) * | 2018-11-23 | 2019-02-26 | 蓝脑科技(厦门)有限公司 | A kind of biofermentation preparation method of the active peptides with white-skinned face function |
| CN110664742A (en) * | 2019-10-29 | 2020-01-10 | 易金阳 | Eye drops for relieving asthenopia and preparation method and application thereof |
| CN112625089A (en) * | 2020-12-14 | 2021-04-09 | 四川省恩乐生物工程有限公司 | Active protein peptide gene with excellent repairing effect and coded protein peptide |
| CN112625089B (en) * | 2020-12-14 | 2023-01-24 | 四川省恩乐生物工程有限公司 | Active protein peptide gene with excellent repairing effect and coded protein peptide |
| CN115154614A (en) * | 2022-07-08 | 2022-10-11 | 广西福莱明生物制药有限公司 | Fusion factor formula combination process |
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