CN103755812A - GM-CSF-HER2 recombinant protein, and preparation method and applications thereof - Google Patents
GM-CSF-HER2 recombinant protein, and preparation method and applications thereof Download PDFInfo
- Publication number
- CN103755812A CN103755812A CN201310684698.9A CN201310684698A CN103755812A CN 103755812 A CN103755812 A CN 103755812A CN 201310684698 A CN201310684698 A CN 201310684698A CN 103755812 A CN103755812 A CN 103755812A
- Authority
- CN
- China
- Prior art keywords
- recombinant protein
- csf
- her2
- cell
- preparation
- 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.)
- Pending
Links
Images
Abstract
The invention relates to a GM-CSF-HER2 recombinant protein, and a preparation method and applications thereof, and specifically relates to a GM-CSF-HER2 recombinant protein which can be used for preparation of antitumor drugs. Amino acid sequence of the GM-CSF-HER2 recombinant protein is represented by SEQ ID No.1. The preparation method comprises following steps: gene sequences of GM-CSF and HER2 are cloned into plasmid pCEP4 via enzyme digestion for recombinant plasmid construction; human embryo kidney 293 cells are transfected by the obtained recombinant plasmid, and recombinant protein expression is performed; and the obtained recombinant protein is purified and collected. The recombinant protein can be used for producing drugs used for treating cancers such as breast cancer. According to the preparation method, genetic engineering method is adopted; tumor specific antigen, transducin, and GM-CSF which is a key agent for in vitro induction of DC cells are combined; the fusion protein which possesses specificity and is capable of prolonging DC cell life span is prepared; prolonging of DC cell survival time and starting of specific immune response are ensured; the GM-CSF-HER2 recombinant protein is capable of avoiding additional adding of GM-CSF in routine DC cell culturing, and insufficient of GM-CSF in culturing cycles; and coexpression GM-CSF is capable of promoting cytotoxic effects.
Description
[technical field]
The present invention relates to GM-CSF-HER2 recombinant protein and methods and applications thereof, be specifically related to the methods and applications of a kind of GM-CSF-HER2 recombinant protein that can be used in medicine for treating tumor thing and preparation method thereof, making DC vaccine.
[background technology]
One of basic reason of tumor development is that the immunity system of body can not be controlled effectively to its growth, this is corrected, and making the immunity system of body can effectively produce the immunological rejection of tumour cell is the problem that people study always in the last hundred years.Tumor vaccine is the main contents of this aspect research, and in numerous tumor vaccines, dendritic cell (dendritic cells, DC) the knurl seedling that the vital role according to antigen presenting cell in tumour immunity designs is to study at present the most popular direction.Antigen is caught, processes and processed to the immunne response of body first by antigen presenting cell (antigen presenting cell, APC), then by antigen presentation to T, bone-marrow-derived lymphocyte, thereby cause a series of immunne response.Dendritic cell (dendritic cell, DC), as the strongest APC of function, can activate tranquillization type T cell (naive T cell), excites initial immunne response, brings into play in vivo powerful immune surveillance function.DC functional defect in tumour patient body, effectively submission tumour antigen, causes immune incapability or immunological tolerance, and tumour is developed.The most noticeable research work of application DC knurl seedling treatment tumour is that application antigen or antigenic peptide impact sensitization (pulsing) DC in vitro, then by it feedback or immunization to row knurl host, carry out immunotherapy of tumors, proved this therapy can induce significantly body produce Peptide-specific CTL, produce protective immunological reaction and can treat the lotus knurl model of having set up, these researchs provide new thinking for the treatment of tumour, DC is adopted and fed back in the clinical treatment for tumour patient, obtained certain curative effect, show that this therapy has the feasibility of certain clinical application.
The antigen presentation function of DC is the basis of DC knurl seedling anti-tumor immunotherapy, and external a large amount of induction amplifications of DC are prerequisites of DC knurl seedling anti-tumor immunotherapy, and the external structure of DC knurl seedling is the key of the anti-tumor immunotherapy of DC knurl seedling.
Along with the development of the external evoked amplification technique of DC and Protocols in Molecular Biology and genetic engineering technique are applied to the discovery of DC knurl seedling construction process, make the advantages such as output increases, purity improves, preparation is easier, more can meet the needs of experiment and clinical study.DC knurl seedling is more ripe for experiment and the clinical study of immunotherapy of tumors, and DC knurl seedling is expected to become a kind of effective means of immunotherapy of tumors.Therefore the present invention prepares DC vaccine in conjunction with stimulations DC cells such as cytokines and can effectively excite antigen specific immune in body to react to build tumour specific antigen, and for prevention and the treatment of tumour.
Foreign literature Sipuleucel-T, Celestia S.Higano et al., nature reviews drug discovery, the 9th volume, 513rd～514 pages, 20100731 disclose a kind of autogenous cell immunotherapy, a kind of preparation method who produces tumor of prostate is wherein disclosed, wherein GM-CSF is obtained to fusion rotein with prostatic acid phosphatase (PAP) restructuring that can be used as therapeutic prostate cancer vaccine targets, activate DC, produce the tumor vaccine of prostatic cancer specific.
HER2, proto-oncogene ErbB-2 (human epidermalgrowth factor receptor-2, HER2) gene, has tyrosine kinase activity, is that important breast cancer diagnosis and prognosis judges the factor.Crossing of HER2 albumen expressed, and makes its clinical characters and biological behaviour have Special Manifestations, and its treatment pattern also makes a big difference with the mammary cancer of other types, and therefore HER2 is positive or cross the mammary cancer of expressing and be different from the mammary cancer of other types.
[summary of the invention]
The DC cell of inducing for present stage application ordinary method has the deficiency of the aspects such as the not high and too early apoptosis of antigen-specific.The invention provides a kind of preparation method of GM-CSF-HER2 recombinant protein, the DC cell of induction is all greatly increased, for follow-up specific immune response provides good basic condition in antigen-specific with on life cycle.The present invention is that immunotherapy of tumors is had laid a good foundation.
For achieving the above object, design a kind of GM-CSF-HER2 recombinant protein, its aminoacid sequence is as shown in SEQ ID NO:1.
Prepare a method for above-mentioned recombinant protein, it is characterized in that the method is comprised of following steps:
A. by the gene order of the gene order of GM-CSF and HER2 by enzyme cutting clone to construction recombination plasmid in plasmid pCEP4,
B. Transfected Recombinant Plasmid human embryonic kidney 293 cell carries out the expression of recombinant protein,
C. purifying is collected described recombinant protein.
The fusion of GM-CSF and HER2 being carried out to recombinant protein has feasibility, and object product can carry out the preparation of DC vaccine, therefore the gene fragment of 2 directly synthetic albumen is linked to above carrier by restriction enzyme site, by applicable expression system, carry out again the expression of albumen, whole process has related to gene molecule clone recombinant technology, protein expression and purification technique, technological method used all conforms with the technological method that this field relates to; Because GM-CSF-HER2 recombinant protein belongs to external source recombinant protein, the expression vector of selecting and expression system are the expression that pCEP4 and human embryonic kidney cell 293 are all applicable to external source recombinant protein, select correct expression vector and expression system can improve success ratio and the expression amount of expression of recombinant proteins, be also convenient to follow-up protein purification.
Described enzyme is cut to: position, enzyme point of contact Hind III and Not I are added in the gene order two ends at GM-CSF, at the gene order two ends of HER2, adds position, enzyme point of contact Xho I and BamH I.The selection of restriction enzyme site need be according to experiment purpose, the multiple clone site that comprises for selected carrier in conjunction with the characteristic of exogenous genetic fragment and select to be beneficial to enzyme and cut the site of combination.At GM-CSF gene order two ends, add restriction enzyme site and should be Hind III and Not I and add restriction enzyme site Xho I and BamH I at HER2 gene order two ends, can be beneficial to structure and the expression of recombinant protein gene.
By Ni-NTA post, carry out described separation and purification.Because GM-CSF-HER2 recombinant protein is with 6His label, and need to separation and purification under non-Denaturing, so select Ni-NTA column separating purification.Selecting Ni-NTA post is also to determine according to the object after recombinant protein self-characteristic and purifying and usability.
Described recombinant protein can be as the medicine of production for treating tumour.
Described tumour is mammary cancer.
Described recombinant protein is prepared a method for DC vaccine, and it is characterized in that stimulates the DC cell of inducing healthy C57BL/6 mouse to prepare via marrow with described recombinant protein, makes DC vaccine.DC cell can be formed by peripheral blood mononuclear cell (PBMC) induction, but proportion is less, be beneficial to the required peripheral blood of experimentation on animals still less, so be unfavorable for the acquisition of DC, select healthy BALB/C mice bones of limbs marrow to be prepared into single cell suspension, add the external evoked mouse DC of going out of cytokine cell, utilize GM-CSF-HER2 recombinant protein to activate out DC vaccine compared with the induction of ten-strike.
The concentration that is used for the recombinant protein that stimulates induction is 25～100ug/ml.
The concentration that is used for the recombinant protein that stimulates induction is 25～50ug/ml.
Compared with the method for utilizing tumor cell lysate induction DC cell with routine, the present invention has the following advantages:
(1) utilize gene engineering method by the key reagents GM-CSF combination of tumour specific antigen and transducer and external evoked DC cell, prepare the fusion rotein that there is specificity and can delay DC cell survival, guaranteed the prolongation of DC cell survival time and the startup of specific immune response;
(2) the successful preparation of GM-CSF-HER2 recombinant protein, has avoided the extra interpolation of GM-CSF in conventional DC cell cultures, solved the shortcoming of GM-CSF deficiency in culture cycle, and coexpression GM-CSF also can promote cytotoxicity;
(3) strengthened the specificity of tumour antigen, solved targetedly the immunotherapy of tumors take HER2 as significant antigen, for clinical therapy of tumor provides new foundation.
[accompanying drawing explanation]
Fig. 1 is that different boosting vaccine body ELISPOT detect IFN-gama spot number;
Fig. 2 is that different boosting vaccine body specific reaction ELISPOT detect IFN-gama spot;
Fig. 3 is volume of tumor mass comparison diagram on the same group not;
Fig. 4 is that various dose recombinant protein stimulates body specific reaction ELISPOT to detect analysis chart;
Fig. 5 is 4T1-HER2 cellulotoxic experiment comparison diagram;
Fig. 6 is 4T1 cellulotoxic experiment comparison diagram;
Fig. 7 is plasmid pCEP4 collection of illustrative plates.
Details as Follows for the collection of illustrative plates of Fig. 7:
CMV promotor: 1-588
Multiple clone site: 619-676
SV40 polyadenylation signal site: 685-926
Replication origin: 1344-3319
EBNA-1 gene (complementary strand) site: 3620-5545
Ampicillin resistance gene site: 6171-7031
Replication origin: 7040-7815
TK promotor: 8183-8345
Hygromycin gene site: 8409-9419
TK polyadenylation signal site: 9431-9702
[embodiment]
In order to make object of the present invention, technical scheme and advantage clearer, the present invention is further elaborated.Production unit in the application is all the common equipment of this area, should be appreciated that specific embodiment described herein, only in order to explain the present invention, is not intended to limit the present invention.
Embodiment 1:
The special encoding sequence of human breast cancer cell SKBr3 specific antigens HER2 and granular leukocyte colony stimulating growth factor GM-CSF encoding sequence are recombinated.
Wherein, GM-CSF is from NCBI for the granular leukocyte colony stimulating growth factor, and its sequence number is AAA52578, and its encoding sequence is:
tqpwehvnai qearrllnls rdtaaemnet vevisemfdl qeptclqtrl elykqglrgs 60
ltklkgpltm mashykqhcp ptpetscatq iitfesfken lkdfllvipf dc 112
GM-CSF gene order two ends add restriction enzyme site Hind III and Not I;
Human breast cancer cell SKBr3 specific antigens HER2 is from NCBI, and its sequence number is NP_004439, and its encoding sequence is:
gcpaeqrasp ltsiisavvg illvvvlgvv fgilikrrqq kirk 44
At HER2 gene order two ends, add restriction enzyme site Xho I and BamH I;
Utilize gene recombination technology that above-mentioned two fragment gene fragments are arrived to plasmid pCEP4 construction recombination plasmid by enzyme cutting clone, plasmid pCEP4 collection of illustrative plates as shown in Figure 7; By recombinant plasmid pCEP4 transfection human embryonic kidney 293 cell, carry out the expression of GM-CSF-HER2 recombinant protein; Then utilize Ni-NTA column separating purification GM-CSF-HER2 recombinant protein;
The mouse mastopathy cell strain 4T1 of construction expression people source HER2, i.e. 4T1-HER2 cell strain, and use RPMI-1640 substratum, add the NaHCO of 1.5g/L
3, the glucose of 2.5g/L, the Sodium.alpha.-ketopropionate of 0.11g/L, 10% heat-inactivated foetal calf serum, 100U/ml penicillin, 100ug/ml Streptomycin sulphate is cultivated;
Choose BALB/C mice, build 4T1-HER2 animal model;
Get healthy BALB/C mice bones of limbs marrow and be prepared into single cell suspension, remove red corpuscle, add perfect medium, cytokine, external evoked go out mouse DC cell, add again the GM-CSF-HER2 recombinant protein having made, through induction, activate and prepare the DC vaccine that GM-CSF-HER2 recombinant protein activates.
(i) the contrast of ordinary method and GM-CSF-HER2 recombinant protein induction DC excitating organism immune response ability:
Ordinary method refers to, utilizes tumor cell line lysate to remove to induce DC cell.
HER2 is proto-oncogene ErbB-2 (human epidermalgrowth factor receptor-2, HER2) gene, there is tyrosine kinase activity, HER2 is that important breast cancer diagnosis and prognosis judges the factor, crossing of HER2 albumen expressed, make its clinical characters and biological behaviour have Special Manifestations, its treatment pattern also makes a big difference with the mammary cancer of other types, and therefore HER2 mammary cancer positive or that excessively express is different from the mammary cancer of other types.DC vaccine prepared by the recombinant protein of granulocyte-macrophage colony stimutaing factor GM-CSF and mammary cancer vaccine targets HER2 fusion feeds back BALB/C mice, can make T cell and CD4+ and the CD8+T cell generation IFN-gama of mouse, and higher than simple HER2 albumen; Utilize GM-CSF-HER2 recombinant protein to carry out the test of 4T1-HER2 breast cancer animal model, result shows the growth that can obviously suppress tumour with blank, physiological saline contrast GM-CSF-HER2 recombinant protein, and inhibition is relevant with protein concentration.
The DC vaccine of getting two kinds of method inductions feeds back BALB/C mice, after utilizing enzyme linked immunological spotting method (ELISPOT) detection vaccine to feed back, T cell produces the amount of IFN-gama, and the amount of CD4+ and CD8+T cell response generation IFN-gama goes to assess the ability of body generation specific immune response.
The different boosting vaccine body of table one ELISPOT detects IFN-gama spot number
| Group | Spot number (individual) |
| |
35 |
| Conventional method (HER2) | 169 |
| Recombinant protein (GM-CSF-HER2) | 572 |
The different boosting vaccine body of table two specific reaction ELISPOT detects IFN-gama spot number
Fig. 2 data presentation, GM-CSF-HER2 recombinant protein stimulates body to produce the ability of HER2 specific immune response will be higher than ordinary method, and difference between the two has statistical significance (P < 0.05).
(ii) GM-CSF-HER2 recombinant protein optimal dose
4T1-HER2 cell strain is to be built and form through Retroviral Transfer mouse breast cancer 4T1 cell strain by the cDNA of carrier source HER2.
Choose BALB/C mice and build 4T1-HER2 breast cancer animal model, according to blank, physiological saline, fusion rotein concentration, be that five groups of 25ug/ml, 50ug/ml, 100ug/ml inject respectively mouse, every group 10, within every 5 days, measure and calculate volume of tumor mass size (mm
3) V=(a*b
2)/2, the diameter of the big or small central shaft that a, b are tumor mass.
Not volume of tumor mass on the same group of table three
In conjunction with upper table and Fig. 3 data presentation, GM-CSF-HER2 recombinant protein can obviously suppress the growth of tumour, and inhibition is relevant with injected dose, and between there is statistical significance (P < 0.05).
After utilizing enzyme linked immunological spotting method (ELISPOT) detection to inject respectively mouse by above five groups, make T cell produce the amount of IFN-gama.According to the data of Fig. 4 demonstration, show, low dosage GM-CSF-HER2 recombinant protein just can be induced body generation specific immune response.
(iii) cellulotoxic experiment
Respectively using 4T1-HER2 and 4T1 cell as target cell.Get respectively injection conventional albumen and the BALB/C mice splenocyte of GM-CSF-HER2 recombinant protein after 7 days, after 4% paraformaldehyde is hatched with action effect cell.According to difference effect target than by cell co-cultivation, every group do three parallel, after 5 days, utilize chromium method for releasing detection specificity cytotoxicity.
Table four 4T1 cellulotoxic experiment
Table five 4T1-HER2 cellulotoxic experiment
Fig. 5 data presentation, GM-CSF-HER2 recombinant protein kills and wounds the effect of expression specificity HER2 cell will be higher than conventional method, there is statistical significance between the two, and the otherness between different effect target ratio also has statistical significance (P < 0.05).
By Fig. 6 data presentation, show that conventional method and GM-CSF-HER2 recombinant protein all can kill and wound mammary cancer 4T1 cell, no significant difference between the two, and different effect target than between also no significant difference.
Claims (9)
1. a GM-CSF-HER2 recombinant protein, its aminoacid sequence is as shown in SEQ ID NO:1.
2. a method of preparing recombinant protein described in claim 1, is characterized in that the method is comprised of following steps:
A. by the gene order of the gene order of GM-CSF and HER2 by enzyme cutting clone to construction recombination plasmid in plasmid pCEP4,
B. Transfected Recombinant Plasmid human embryonic kidney 293 cell carries out the expression of recombinant protein,
C. purifying is collected described recombinant protein.
3. the method for preparing recombinant protein as claimed in claim 2, is characterized in that described enzyme is cut to: position, enzyme point of contact Hind III and Not I are added in the gene order two ends at GM-CSF, at the gene order two ends of HER2, adds position, enzyme point of contact Xho I and BamH I.
4. the method for preparing recombinant protein as claimed in claim 2, is characterized in that carrying out described separation and purification by Ni-NTA post.
Described in a claim 1 recombinant protein for the application of anti-tumor medicine.
6. recombinant protein as claimed in claim 5 is used for the treatment of the application of tumour medicine, it is characterized in that described tumour is mammary cancer.
7. described in claim 1, recombinant protein is prepared the method for DC vaccine, it is characterized in that the DC cell that stimulates the healthy C57BL/6 mouse of induction to prepare via marrow with described recombinant protein making DC vaccine.
8. recombinant protein is prepared the method for DC vaccine as claimed in claim 7, and the concentration that it is characterized in that the recombinant protein for stimulating induction is 25～100ug/ml.
9. recombinant protein is prepared the method for DC vaccine as claimed in claim 7, and the concentration that it is characterized in that the recombinant protein for stimulating induction is 25～50ug/ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310684698.9A CN103755812A (en) | 2012-12-14 | 2013-12-13 | GM-CSF-HER2 recombinant protein, and preparation method and applications thereof |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210544431.5 | 2012-12-14 | ||
| CN2012105444315A CN102993312A (en) | 2012-12-14 | 2012-12-14 | GM-CSF-HER2 recombinant protein, method and application thereof |
| CN201310684698.9A CN103755812A (en) | 2012-12-14 | 2013-12-13 | GM-CSF-HER2 recombinant protein, and preparation method and applications thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103755812A true CN103755812A (en) | 2014-04-30 |
Family
ID=47922471
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012105444315A Pending CN102993312A (en) | 2012-12-14 | 2012-12-14 | GM-CSF-HER2 recombinant protein, method and application thereof |
| CN201310684698.9A Pending CN103755812A (en) | 2012-12-14 | 2013-12-13 | GM-CSF-HER2 recombinant protein, and preparation method and applications thereof |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012105444315A Pending CN102993312A (en) | 2012-12-14 | 2012-12-14 | GM-CSF-HER2 recombinant protein, method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN102993312A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774270A (en) * | 2015-05-06 | 2015-07-15 | 河北利同康生物科技有限公司 | Adenocarcinoma specificity EpCAM-GM-CSF genetic recombinant fusion protein and preparation method thereof |
| WO2019062790A1 (en) * | 2017-09-29 | 2019-04-04 | Lan Keng Li | Novel conjugates and uses thereof |
| WO2023011662A1 (en) * | 2021-08-06 | 2023-02-09 | 上海赛金生物医药有限公司 | Anti-her-2 antibody-granulocyte regulatory factor fusion protein, preparation method therefor and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111303302A (en) * | 2020-03-19 | 2020-06-19 | 芜湖天明生物技术有限公司 | Soluble efficiently-expressed rChGM-CSF-IFN α fusion protein and preparation method and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101638440A (en) * | 2008-07-29 | 2010-02-03 | 中国人民解放军军事医学科学院基础医学研究所 | Heterogenetic antigen-Fc fusion protein capable of inducing antitumor immunity of organism and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101966335B (en) * | 2009-07-27 | 2013-02-06 | 湖南惠霖生命科技有限公司 | Novel HER2/neu gene-modified dendritic cell vaccine |
-
2012
- 2012-12-14 CN CN2012105444315A patent/CN102993312A/en active Pending
-
2013
- 2013-12-13 CN CN201310684698.9A patent/CN103755812A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101638440A (en) * | 2008-07-29 | 2010-02-03 | 中国人民解放军军事医学科学院基础医学研究所 | Heterogenetic antigen-Fc fusion protein capable of inducing antitumor immunity of organism and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| CELESTIA S.HIGANO ET AL.: "Sipuleucel-T", 《NATURE REVIEWS DRUG DISCOVERY》, vol. 9, 3 July 2010 (2010-07-03) * |
| 唐三元 等: "HER2/neu和GM-CSF共转染树突状细胞的免疫杀伤研究", 《细胞与分子免疫学杂志》, vol. 24, no. 1, 31 December 2008 (2008-12-31), pages 76 - 79 * |
| 赵丽丽 等: "HER-2/neu肿瘤疫苗的研究进展", 《中国肿瘤生物治疗杂志》, no. 02, 3 April 2010 (2010-04-03) * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104774270A (en) * | 2015-05-06 | 2015-07-15 | 河北利同康生物科技有限公司 | Adenocarcinoma specificity EpCAM-GM-CSF genetic recombinant fusion protein and preparation method thereof |
| CN104774270B (en) * | 2015-05-06 | 2017-08-08 | 河北利同康生物科技有限公司 | A kind of gland cancer specificity EpCAM GM CSF gene recombinant fusion proteins and preparation method thereof |
| WO2019062790A1 (en) * | 2017-09-29 | 2019-04-04 | Lan Keng Li | Novel conjugates and uses thereof |
| WO2023011662A1 (en) * | 2021-08-06 | 2023-02-09 | 上海赛金生物医药有限公司 | Anti-her-2 antibody-granulocyte regulatory factor fusion protein, preparation method therefor and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102993312A (en) | 2013-03-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109554353B (en) | Isolated recombinant oncolytic poxvirus, pharmaceutical compositions and use thereof in a medicament for the treatment of tumors and/or cancers | |
| CN103755812A (en) | GM-CSF-HER2 recombinant protein, and preparation method and applications thereof | |
| CN109055430A (en) | A kind of preparation method for co-expressing IL18 and CCL19 albumen and targeting MUC1 gene C AR-T cell | |
| WO2020259151A1 (en) | Preparation method and application of ctl cell | |
| US11213563B2 (en) | Polypeptide and use thereof | |
| US11466053B2 (en) | Polypeptide and use thereof | |
| CN110214144B (en) | Polypeptides and uses thereof | |
| US11142547B2 (en) | Polypeptide and use thereof | |
| CN103788213B (en) | A kind of GM-CSF-TAT-CEA recombinant protein and methods and applications thereof | |
| US11820836B2 (en) | Polypeptide and use thereof | |
| CN110167956B (en) | Polypeptide and application thereof | |
| CN115850377A (en) | Tumor neoantigen polypeptide based on NRAS gene Q61K mutation and application thereof | |
| US11396528B2 (en) | Polypeptide and use thereof | |
| CN109790224A (en) | Tumor-antigen peptide and its application derived from CACNA1H | |
| CN110072876B (en) | Polypeptide and application thereof | |
| CN110139875B (en) | COL14A 1-derived tumor antigen polypeptide and application thereof | |
| CN116948004B (en) | Tumor new antigen polypeptide aiming at CTNNB1 gene H36P mutation and application thereof | |
| CN100419078C (en) | Expression method of glucosal related protein and application thereof | |
| CN111057690A (en) | Tumor-associated gene BRAF mutation-associated antigen short peptide and application thereof | |
| CN111087448A (en) | Tumor-associated gene JAK2 mutation-associated antigen short peptide and application thereof | |
| CN110845583A (en) | Varicella zoster virus associated antigen short peptide and application thereof | |
| CN103923176A (en) | Oligopeptide with breast cancer resisting activity and application thereof | |
| CN102191271A (en) | Anti-tumor recombinant plasmid as well as gene vaccine, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140430 |
|
| RJ01 | Rejection of invention patent application after publication |