CN108276476B - A kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof - Google Patents
A kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof Download PDFInfo
- Publication number
- CN108276476B CN108276476B CN201711448093.4A CN201711448093A CN108276476B CN 108276476 B CN108276476 B CN 108276476B CN 201711448093 A CN201711448093 A CN 201711448093A CN 108276476 B CN108276476 B CN 108276476B
- Authority
- CN
- China
- Prior art keywords
- cell
- chicken
- targeting peptides
- dendritic cells
- source
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/385—Haptens or antigens, bound to carriers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Immunology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Inorganic Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof.The amino acid sequence of the chicken marrow source Dendritic Cells targeting peptides is as shown in SEQ ID NO.1.The present invention filters out chicken marrow source DC advantage combination polypeptide SP using phage random peptide library and full Cell differentials screening technique, compared with the source of people DC targeting peptides delivered, it can be incorporated into chicken source DC strongly, show good targeting and specificity, therefore targeting peptides SP can be used as guide's molecule of the drugs such as gene targeting carrier, lay the foundation for cell-specific small molecule therapy, or be used in combination with vaccine, bird vaccine Immune efficiency is improved, body is promoted to generate immunity ahead of time.
Description
Technical field
The present invention relates to a kind of chicken marrow source Dendritic Cells targeting peptides and application thereof, belong to biomedicine technical field.
Background technique
The fast development of immunology and molecular biology has pushed the development of new generation vaccine, has utilized immunocyte and vaccine
The interaction of antigen, Lai Youhua body response, to accelerate the speed of immune response, improve the specificity of acquired immunity.Closely
Nian Lai, magnetic target therapy and prevention have become technical field of pharmaceutical biotechnology research hotspot, have been applied to the directions such as human tumour
Treatment, wherein drug can be effectively delivered into target site by the biological immune therapy of targeting Dendritic Cells, medicament is used in reduction
Amount and administration number of times improve the therapeutic effect of drug, reduce adverse reaction.Dendritic Cells (dendritic cells, DCs)
Be the strongest professional antigen of function that is currently known in delivery cell, can effectively activate primary tape T cell, in natural immune system and
It plays a significant role in acquired immune system.Researcher utilizes phage display technology linear 12 peptide library technology, and screening has obtained one
Kind human peripheral DCs targeting peptides obtain splendid immune effect by it with after Antigen Fusion expression.In addition, researcher is sharp
With 7 peptide library technology of bacteriophage ring, mouse bone marrow cells source DC targeting peptides have been filtered out, have been made with the chitosan nano particle of antigen coupling
For vaccine, Immune efficiency is significantly improved.
Since variant or the appearance of virulence enhancing strain, the effect of existing vaccine product are increasingly subject to influence, new product
Develop change is how to improve vaccine potency, such as improves the intensity of protective immune response, generates immunity ahead of time or prolong
Long duration of immunity etc., targeting vaccine become new generation vaccine research emphasis.The relevant research of poultry immunity lags behind mammal, shadow
The field of the development and exploitation of its newtype drug of sound and biological products, fowl magnetic target therapy and prevention is even more to rarely have to be related to.Although
Existing mammal DC targeting peptides can and the DC of multiple source of species different degrees of combination, but the combination to fowl source DC occurs
Effect is poor.Bird vaccine is not yet related to target administration, has not yet to see the report in relation to fowl source DC targeting peptides.
The present invention is that crucial this feature of antigen presenting cell is set out according to Dendritic Cells, and design, which filters out, to be directed to
The good linear targeting peptides of chicken Dendritic Cells compatibility, development and exploitation to the novel DC targeting vaccine of poultry have weight
Want meaning.The present invention is separately cultured condition to chicken marrow DCs and gropes, and identifies its form, phenotype, function, is intended to
The higher cell of purity target cell as a filter is obtained, utilizes 12 peptide library technology of phage display, then with chicken marrow source DC
For target cell, using bone marrow cell as negative selection cell, it is contemplated that screening obtains excellent linear 12 peptide in conjunction with chicken DCs, and leads to
Crossing the experimental verifications such as Phage-ELISA, flow cytometry, immunofluorescence, it combines effect, can target chicken source DC to obtain
Small peptide, can be used as the drug leads molecule such as gene targeting carrier, lay the foundation for cell-specific small molecule therapy.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of chicken marrow source Dendritic Cells targeting peptides and application thereof.
In order to achieve the above object, present invention employs following technological means:
A kind of chicken marrow source Dendritic Cells targeting peptides of the invention, are named as SP, amino acid sequence such as SEQ
Shown in IDNO.1.
The nucleotide sequence of the coding chicken marrow source Dendritic Cells targeting peptides contains the nucleotide sequence
Expression vector and host cell containing the expression vector are also within protection scope of the present invention.
Further, the invention also provides the chicken marrow source Dendritic Cells targeting peptides to prepare chicken marrow source tree
Purposes in prominent shape cell-targeting drug.
Wherein, it is preferred that the drug is vaccine.
Compared to the prior art, the beneficial effects of the present invention are:
1, the present invention is combined using phage random peptide library and full Cell differentials screening technique screening chicken marrow source DC advantage
Polypeptide provides material base to target vaccine for research and development chicken source DC.One breeder source DC targeting peptides SP of the present invention
(shown in SPHLHTSSPWER, SEQ ID NO.1) can be incorporated into Ji Yuan compared with the source of people DC targeting peptides delivered strongly
DC, it is shown that good targeting and specificity.
2, the present invention filters out chicken marrow source DC advantage knot using phage random peptide library and full Cell differentials screening technique
Close polypeptide SP (shown in SPHLHTSSPWER, SEQ ID NO.1) have immunogenicity is low, relative molecular mass is small, in blood and
Remove that fast, tissue penetration is strong, high with receptor affinity and can gulp down by receptor mediated endocytosis formation in non-target tissue
Bite corpusculum and enter the features such as intracellular, and be readily synthesized, can be reduced by structural modification its degradation rate in vivo and
Its side effect.Targeting small peptide can be used in combination with a variety of drug conveying carriers, reach good Targeting Effect, the effect of acquisition
It is that plain vaccine is not achieved.
3, the present invention obtains a chicken marrow source DC targeting peptides SP through subtractive screen using 12 peptide library technology of phage display
(shown in SPHLHTSSPWER, SEQ ID NO.1), it is thin which, which can be used as the drug leads molecule such as gene targeting carrier,
The treatment of born of the same parents' specific small molecule lays the foundation, or is used in combination with vaccine, improves bird vaccine Immune efficiency, and body is promoted to do sth. in advance
Generate immunity.
Detailed description of the invention
Fig. 1 is chicken marrow source Dendritic Cells optical microphotograph sem observation result;
A is the 6th day om observation result of cell culture;B is cell om observation result after LPS is stimulated for 24 hours;
Fig. 2 is the expression of chicken marrow source DC surface molecular mark after cultivating 7d;
Fig. 3 is mixed lymphocyte reaction (MLP);
Fig. 4 is sequencing comparison result and duplicate sequence results after fourth round screening;
Fig. 5 is the flow cytometer showed result that FITC marks peptide and the affine effect of chicken source DC;
Fig. 6 is fluorescence microscope fluorescent peptide effect in conjunction with DC;
Fig. 7 is that laser co-focusing observes fluorescent peptide binding site;
Fig. 8 is Competitive assays ELISA result;
Fig. 9 is cell factor IFN-γ and IL-10 transcriptional level testing result.
Specific embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and
It is apparent.But examples are merely exemplary, and it is not intended to limit the scope of the present invention in any way.Those skilled in the art answer
It should be appreciated that without departing from the spirit and scope of the invention can details to technical solution of the present invention and form repair
Change or replace, but these modifications and replacement are fallen within the protection scope of the present invention.
The Isolation and identification of 1 chicken marrow source Dendritic Cells of embodiment
1, chicken marrow source Dendritic Cells is separately cultured
The chicken of 30 ages in days or so impregnates 10min in 75% ethyl alcohol after putting to death, take out chicken femur and shin under aseptic condition
Bone is drawn with syringe and contains 2% dual anti-PBS (pH 7.2), rinses marrow to pulp cavity and bleaches;Washing lotion is collected, it is dual anti-with containing 2%
PBS is washed 2 times, and each 1000r/min is centrifuged 5-10min, abandons supernatant;It collects precipitating and sedimentation cell is resuspended with PBS, then
Bone marrow cell suspension is slowly added into the centrifuge tube containing Histopaque-1119 in 1:1 ratio, 2500r/min centrifugation
30min;The misty cell of white at intermediate interface is collected, PBS is resuspended, and 1500r/min is centrifuged 10min, abandons supernatant, collects
Precipitating, and cell is resuspended containing 10%FBS and 1% dual anti-RPMI1640 complete medium with 1mL, carry out cell count;Will
To bone marrow cell be inoculated in six porocyte culture plates, every hole adds 3mL to contain chicken rGM-CSF (50ng/mL) and chicken rIL-4
The RPMI1640 complete medium of (50ng/mL), final concentration of the 2 × 10 of cell6Then/mL sets 37 DEG C, 5%CO2Culture
It is cultivated in case;3rd and the 5th day, half amount changed liquid, removed heteroproteose cell and dead cell, while supplying rGM-CSF and rIL-4.Chicken marrow
After cell Fiber differentiation 6d, mature DCs are obtained after the LPS stimulation that whole mass concentration is 200ng/mL.
2, chicken marrow source Dendritic Cells morphologic observation
During cell culture, Application Optics inverted microscope carries out metamorphosis observation to the DC of culture, as a result sees figure
Shown in 1A.After cultivating 2h, cell starts gradually adherent, cell bright color, small volume, and a small amount of distribution is made of several cells
Cell microcolony;After for 24 hours, attached cell slightly increases, and cell colony quantity is still less;3d, attached cell number increase, carefully
Born of the same parents' colony is more, and part cell is still in suspension;5d, cell colony increase, and colony appearance is loose adherent
There are multiple burrs in phenomenon, cell surface;It is visible thin compared with the cell without stimulation under high power lens after LPS stimulation culture for 24 hours
There are many dendroids, furcella shape protrusion in cellular surface, and protrusion is coarse longer, as shown in Figure 1B.
3, the purity and phenotype of flow cytomery chicken marrow source Dendritic Cells
The DC that each group is collected into is respectively charged into centrifuge tube, and 1000r/min is centrifuged 10min, is abandoned supernatant, is used PBS
Cell, adjustment cell concentration to 5 × 10 is resuspended in (0.01mol/L)5/mL.To for identifying that the cell of DC purity (does not stimulate
DC the anti-chicken MHCⅡ antibody of anti-human CD11c or the FITC label of PE label is added in), room temperature, which is protected from light, is incubated for 20min, PBS washing
2 times, 1000r/min is centrifuged 5min, and cell then is resuspended with 200 μ L PBS, is detected using flow cytometer.To for reflecting
Determine the anti-chicken CD40 of mouse or the anti-chicken CD86 antibody of mouse that unstressed configuration label is added in the cell of DC maturation phenotype, be incubated at room temperature 20min,
PBS is washed 2 times, and 1000r/min is centrifuged 5min, then adds the sheep anti-mouse igg of PE label, and room temperature, which is protected from light, is incubated for 20min, is used
PBS is washed 2 times, and 1000r/min is centrifuged 5min, and DC then is resuspended with 200 μ L PBS, is finally examined using flow cytometer
It surveys.
Immature cell surface C D11c and MHCII molecular marker, is as a result shown in figure after Flow cytometry Fiber differentiation 7d
Shown in 2, the expression quantity of chicken marrow source DC surface C D11c molecule is that the expression quantity of 84.1%, MHCII molecule is 74.7%.It is mature
For DC compared with immature DC, CD40 has raised about 17%.For mature DC compared with immature DC, CD86 has raised about 11%.
4, Allogeneic Mixed Lymphocyte reacts
React whether identification maturation DC has the function of activated lymphocyte proliferation using Allogeneic Mixed Lymphocyte,
The specific method is as follows: the sucking of 30 age in days chicken anesthetic is lethal, sterile to take its spleen, and it is unicellular that the acquisition of 200 mesh screens is crossed after grinding
Suspension, through the isolated lymphocyte of lymphocyte separation medium, as reacting cells;It is 50 μ g/mL's that whole mass concentration, which is added,
Mitomycin C handles DC, and as stimulation cell, the concentration for adjusting lymphocyte is 2 × 10696 well culture plates are added, often in/mL
100 μ L of hole;To stimulate cell and reacting cells ratio is respectively that 1:1,1:10,1:100 are added DC Jing Guo different disposal and (are divided into
LPS stimulation group and DC group without stimulation), while independent T cell hole is set as negative control and cell-free blank control,
3 repetitions of every hole cell, final volume are 200 μ L, are placed in 37 DEG C, 5%CO2Incubator in cultivate 72h;20 μ L are added in every hole
CCK-8 solution, is incubated for 2h, and microplate reader reads OD450 value.Stimulus index of the DC to Allogeneic Mixed Lymphocyte
(stimulation index, SI) calculation formula are as follows: SI=(ODDC-ODBlank control)/(ODNegative hole-ODBlank control).As a result see Fig. 3, pass through
The mature DC stimulation Allogenic Lymphocytes proliferative capacity of LPS stimulation is significantly higher than DC (the * p < mature without stimulation
0.05, * * p < 0.01), stimulation ability is directly proportional to the concentration of DC.
The Preliminary detection of 2. phage display of embodiment, 12 peptide library subtractive screen targeting peptides and its affine DC ability
1,12 peptide library subtractive screen targeting peptides of phage display
Screening process is divided into four-wheel, specific as follows:
The first round: by 2 × 1011Library phage display 12 peptide library (PhD-12) of pfu is added to the chicken that embodiment 1 obtains
Bone marrow Dendritic Cells (5 × 105/ mL) in, and 30min is incubated at 4 DEG C, by cell precipitation after 2000rpm centrifugation 3min
It is resuspended in the PBS containing 1% bovine serum albumin(BSA) (BSA) and 0.05% polysorbas20, washes repeatedly 3 times;It is slow that 1mL is added
50 μ gently are added after mixed 10min at room temperature in fliud flushing (0.2mol/L glycine-HCI, pH=2.2,1mg/mL bovine serum albumin(BSA))
L Tris-HCl (pH value 9.1) is neutralized;Centrifugation, supernatant are film surface combination bacteriophage elution liquid, eluent respectively with
E.coli ER2738 culture solution (A600nm ≈ 0.5) acutely shakes 4.5h in conical flask, heavy using polyethylene glycol NaCl bis- times
Shallow lake method prepares Phage amplification liquid.
The screening of second~four-wheel: liquid will be expanded the first round and is added to 5 × 1054 DEG C of (negative selection cell) in/mL bone marrow cell
Supernatant is transferred to BMDC after lower incubation 30min, 2000rpm centrifugation 3min, repeats first round step;Third round is by incubation time
It is shorten to 20min, incubation time is shorten to 10min by fourth round.Each wheel screening eluent and amplification liquid use LB isopropyl-sulphur
Generation chloro- 3- indoles-β-thiogalactoside (Xgal) Tet plate culture assay phagocytosis of the bromo- 4- of-β-D- galactoside (IPTG) 5-
Body titre;After 4th wheel biological elimination, 168 plaques are selected at random, are extracted M13 phage single-chain DNA with kit, are surveyed
Sequence.
Number of screening round and input amount, yield are shown in Table 1, after every wheel screening rinses the bacteriophage of non-specific binding,
To expanding after being incorporated into the bacteriophage elution on surface, it is carried out continuously four-wheel elutriation, the rate of recovery is risen in the second wheel, the
Three, after reducing incubation time, the rate of recovery tends to be steady four-wheel.In fourth round Sequencing of plaques result, 168 are sequenced altogether, weight
Multiple rate and sequence results as shown in Figure 4, after four-wheel elutriation, show SP small peptide (SPHLHTSSPWER, SEQ ID NO.1 institute
Show) bacteriophage frequency of occurrences highest, indicate that it may be and the preferable peptide sequence of chicken marrow source DC compatibility.
1 number of screening round of table, input amount and yield
2. the synthesis of polypeptide
It selects the highest peptide SP of repetitive rate (shown in SPHLHTSSPWER, SEQ ID NO.1) in fourth round screening, repeat one
Secondary peptide AM (AMEKYGQQIATQ) and human peripheral DC targeting peptides FY (FYPSYHSTPQRP) is synthesized using Fmoc method, is used in combination
FITC marks small peptide, carries out HPLC purifying (purity > 95%) and mass spectral analysis (completion of Wuhan Xing Hao Pharmaceuticals Ltd).Fluorescence
Peptide powder is dissolved in DMSO in advance, then plus PBS to DMSO final concentration of 30%, -20 DEG C save backup.
3. the detection of small peptide affinity
The affinity of 3.1 Flow cytometry peptides and chicken marrow source DC
12 peptide SP, FY and AM that 25 μ g FITC are marked are added separately to (cell concentration about 10 in the DC of culture the 6th day6/
ML), after 4 DEG C of effect 20min, 1800r/min centrifugations, precipitating is washed twice with PBS, and cell is resuspended with 500 μ lPBS, thin through streaming
Born of the same parents' instrument detects fluorescence signal, and each sample is in triplicate.Testing result as shown in Figure 5, under the same conditions, believe by SP small peptide fluorescence
Number intensity is significantly higher than AM small peptide and source of people DC targeting peptides FY, and it is special to show that it can preferably occur with chicken source Dendritic Cells
Property combine.
3.2 fluorescence microscope fluorescent peptides effect in conjunction with the DC of chicken marrow source
Sterile creep plate is laid in 24 orifice plates, 0.03mg/ml poly-D-lysine, every 200 μ l of hole, room temperature effect is added
30min, abandons liquid, and PBS is washed and will be cultivated the 6th day DC afterwards twice (cell number is about 105/ mL), 50 μ g FITC label 12 peptide SP
It is added separately in each hole with AM, room temperature acts on 30min, and PBS is rinsed three times, fluorescence microscope fluorescent peptide combination effect.
As a result as shown in Figure 6, the fluorescence intensity of SP small peptide and cell combination is higher than the fluorescence intensity of AM small peptide, shows that SP fluorescent peptide can
It is preferably combined with the generation of DC cell.
3.3 laser co-focusings observe fluorescent peptide and chicken marrow source DC binding site
Cellular processes and fluorescent peptide action time and etc. ibid (3.2).After PBS rinsing three times, DIL dyestuff is added
(1:1000 dilution), 4 DEG C of incubation 30min, PBS are rinsed 3 times, pass through the position of confocal laser scanning microscope fluorescent peptide combination DC
It sets.As a result as shown in Figure 7, SP small peptide can preferably be incorporated into the cell surface (yellow that red in Merge and green combination is formed
As common location position-cell membrane), part of fluorescent peptide has entered cell interior (green is rendered as in Merge), identical item
Under part, the fluorescence intensity of AM small peptide combination cell is weaker, in conjunction with ineffective.
3.4 Competitive assays ELISA experiment
Whether the combination that small peptide and DC are verified by Competitive assays ELISA method is in dose dependent, and the specific method is as follows:
96 orifice plates are added in 0.03mg/ml poly-D-lysine, and every 100 μ l of hole after room temperature acts on 30min, abandons liquid, and PBS is washed twice, will be cultivated
The full cell of chicken marrow to the 6th day chicken marrow source DC and newly separated is inoculated in 96 orifice plates (every hole 10 respectively5), 37 DEG C of effects
2h;Then (0 μ g, 25 μ g, 50 μ g, 75 μ g, 100 μ g, 125 μ g, 150 μ g are mended with PBS to 100 μ with various concentration gradient respectively
L) 4 DEG C of SP peptide effect 30min, PBST washing 3 times, are added SP bacteriophage (1011Pfu) in 4 DEG C of incubation 30min;PBST washing 3
After secondary, it is incubated for 10min with the PBS containing 0.1%BSA, the anti-M13 monoclonal antibody (1:1500) of 100 μ L mouse, incubation at room temperature is added
1h;The mountain sheep anti-mouse igg (1:5000) of HRP label, every 100 μ l of hole are added after PBST washing, room temperature acts on 1h, and every hole is added
The TMB developing solution (2 μ l of A liquid 2.5ml, B liquid 2.5ml, C liquid) of 100 μ l, 37 DEG C of incubation 15min, then every hole is added 50 μ L and terminates
Liquid 2mol/L H2SO4, ELISA instrument measures absorbance at each hole 490nm.It is repeated 3 times, and cell negative control is set.
As a result as shown in Figure 8, SP inhibits the combination of phage clone and DC, increases with SP concentration, phage clone is in conjunction with DC
OD490 value is gradually reduced.
Influence of the 3.5 targeting peptides SP to cell IFN-γ and IL-10 transcriptional level
For DC under environmental stimuli, the secretion of cell factor may be subjected to influence.Therefore, using fluorescence quantifying PCR method
The transcriptional level of two key cytokines (IFN-γ and IL-10) after targeting peptides SP and DC is acted on is detected, specific method is such as
Under: the targeting peptides SP and about 5 × 10 that 25 μ g are synthesized6A immature DC collects cell, does not stimulate after 4 DEG C of effect 30min
Cell be used for control cell, by RNA extracts kit specification extraction RNA, reverse transcription is at cDNA.SYBR Green real-
Time RT-PCR: by 2 × SYBR Green PCRMasterMix, template cDNA respectively with primer β -1/ β -2, IFN-1/IFN-
2 and IL10-1/IL10-2 mixes (primer sequence is shown in Table 2), is dispensed into each PCR pipe after mixing, 3 weights of each sample
It is multiple.Real time PCR amplification program are as follows: 95 DEG C of 60s, 95 DEG C of 15s, 60 DEG C of 30s, totally 40 recycle.β-actin is reference gene, phase
Fluorescent quantitation (Relative quantity) is calculated as follows:△△Ct=(CtTarget gene-CtReference gene)Experimental group-(CtTarget gene-
CtReference gene)Control group.Testing result as shown in Figure 9, targeting peptides SP and DC effect after, cell factor IFN-γ and IL-10 transcriptional level
Variation is not significant, shows that the function of DC is not affected by influence.
2 primer sequence of table
To sum up, the present invention is higher by one repeatability of four-wheel subtractive screen acquisition using 12 peptide library technology of phage display
Linear 12 peptide SP (SPHLHTSSPWER), by experiment confirm the peptide to chicken marrow source DC have stronger affinity, display
Good targeting.
Sequence table
<110>Northeast Agricultural University
<120>a kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof
<130> KLPI171047
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 12
<212> PRT
<213> SP
<400> 1
Ser Pro His Leu His Thr Ser Ser Pro Trp Glu Arg
1 5 10
Claims (6)
1. a kind of chicken marrow source Dendritic Cells targeting peptides, are named as SP, which is characterized in that the chicken marrow source dendron shape is thin
The amino acid sequence of born of the same parents' targeting peptides is as shown in SEQ ID NO.1.
2. encoding the polynucleotides of chicken marrow source Dendritic Cells targeting peptides described in claim 1.
3. a kind of expression vector, which is characterized in that contain polynucleotides as claimed in claim 2.
4. a kind of host cell, which is characterized in that contain expression vector as claimed in claim 3.
5. chicken marrow source Dendritic Cells targeting peptides described in claim 1 are preparing chicken marrow source Dendritic Cells targeted drug
In purposes.
6. purposes as claimed in claim 5, which is characterized in that the drug is vaccine.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201711448093.4A CN108276476B (en) | 2017-12-27 | 2017-12-27 | A kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201711448093.4A CN108276476B (en) | 2017-12-27 | 2017-12-27 | A kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN108276476A CN108276476A (en) | 2018-07-13 |
CN108276476B true CN108276476B (en) | 2019-11-22 |
Family
ID=62802400
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201711448093.4A Active CN108276476B (en) | 2017-12-27 | 2017-12-27 | A kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108276476B (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111533785B (en) * | 2020-02-11 | 2022-03-08 | 北京市肿瘤防治研究所 | Targeting immune checkpoint TIM3 binding peptides and uses thereof |
CN117088943B (en) * | 2023-07-04 | 2024-05-14 | 东北农业大学 | Porcine dendritic cell targeting peptide KC1 and application thereof |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101489589A (en) * | 2006-05-19 | 2009-07-22 | 沃尔特及伊莱萨霍尔医学研究院 | Immunogenic compositions |
CN104387453A (en) * | 2014-12-08 | 2015-03-04 | 深圳市同康生物医药有限公司 | Dendritic cell targeted peptide, coding gene and application |
-
2017
- 2017-12-27 CN CN201711448093.4A patent/CN108276476B/en active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101489589A (en) * | 2006-05-19 | 2009-07-22 | 沃尔特及伊莱萨霍尔医学研究院 | Immunogenic compositions |
CN104387453A (en) * | 2014-12-08 | 2015-03-04 | 深圳市同康生物医药有限公司 | Dendritic cell targeted peptide, coding gene and application |
Non-Patent Citations (1)
Title |
---|
Oral Delivery of Probiotics Expressing Dendritic Cell-Targeting Peptide Fused with Porcine Epidemic Diarrhea Virus COE Antigen: A Promising Vaccine Strategy against PEDV;Xiaona Wang等;《Viruses》;20171025;第9卷(第11期);第1-16页 * |
Also Published As
Publication number | Publication date |
---|---|
CN108276476A (en) | 2018-07-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105177031B (en) | T cell of Chimeric antigen receptor modification and application thereof | |
CN105473731B (en) | The separation of itself cancer antigen-specific CD8+ T cell and enrichment procedure | |
CN109789092A (en) | Antigen presenting cell simulates bracket and its preparation and application | |
CN109575118A (en) | It is used to prepare the polypeptide fragment and DC vaccine of DC vaccine | |
CN103951745A (en) | KDR peptides and vaccines comprising the same | |
CN105903006A (en) | Cancer vaccine composition | |
CN103800898A (en) | Tumor specific killer cell preparation and preparation method thereof | |
CN108276476B (en) | A kind of chicken marrow source Dendritic Cells targeting peptides SP and application thereof | |
WO2018233589A1 (en) | Method for preparing clinical-grade car-t cell preparation by transfecting t cell with minicircle dna | |
CN110305906B (en) | PDL 1-targeted lentiviral vector of CAR chimeric receptor and PDL1-CAR-T cell | |
CN110055269A (en) | People's mesothelin Chimeric antigen receptor, its T cell and its preparation method and application | |
CN110317822A (en) | TROP2 Chimeric antigen receptor, its T cell and its preparation method and application | |
CN107287163B (en) | Express the dendritic cells and application thereof of Chimeric antigen receptor | |
AU2020294154A1 (en) | Mini-combined adjuvants carrier-free nanoparticles and preparation method and application thereof | |
CN109021078B (en) | Dendritic cell-targeting affinity peptide TY peptide and application thereof | |
CN108276475A (en) | A kind of chicken marrow source Dendritic Cells targeting peptides AH and application thereof | |
CN104293815A (en) | Nanometer gene vaccine as well as preparation method and application thereof | |
Xia et al. | Human dendritic cell targeting peptide can be targeted to porcine dendritic cells to improve antigen capture efficiency to stimulate stronger immune response | |
CN105504063B (en) | The antineoplastic amalgamation protein of a kind of alexin-albumin and its preparation and application | |
CN107286246A (en) | Treat BMDC of Chimeric antigen receptor modification of glioma and preparation method thereof | |
CN113429481B (en) | Nano antibody molecule of targeted dendritic cell and application | |
CN105969728A (en) | High-cytotoxic activity CIK cells and application in tumor cellular immunotherapy | |
CN109136182A (en) | A kind of polarization and the method and composition for expanding CD4+T cell and the application in the tumour for curing expression specificity antigen | |
CN114887070A (en) | Spleen-targeted nano-drug | |
CN117088943B (en) | Porcine dendritic cell targeting peptide KC1 and application thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |