CN107557336A - A kind of immunocyte of safety-type Chimeric antigen receptor modification of anti-MUC 16 and its application - Google Patents
A kind of immunocyte of safety-type Chimeric antigen receptor modification of anti-MUC 16 and its application Download PDFInfo
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Abstract
The invention discloses a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, immunocyte is modified to obtain the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 through the safety-type Chimerical receptor of anti-MUC 16, the safety-type Chimeric antigen receptor of anti-MUC 16 includes CD8 leader, MUC16 lands, CD8 hinge area, cross-film stimulus structure domain, CD3 ζ stimulus signals conducting regions and chemical induction element and its application, toxicity is alleviated, improves Clinical efficacy and security.
Description
Technical field
The present invention relates to field of biological genes, is a kind of safety-type Chimeric antigen receptor modification of anti-MUC 16 in particular
Immunocyte and its application.
Background technology
Oophoroma is to cause the fifth-largest tumour of women die.Due to the disguise of its disease locus, and lack effective
Sensitive index, cause oophoroma fatal rate to occupy first of all gynecological tumors, serious threat caused to women life.Most
Recent studies on as shown by data, 90% oophoroma is ovarian epithelial carcinoma (epithelial ovarian cancer, EOC), and is disliked
The property high human ovarian cancer patients' five year survival rate of degree is even below 30%.MUC16, also known as CA125, gene are located at human chromosomal
19p13.2, size 179000bp, it is a kind of high molecular weight glycoproteins for being expressed in all kinds of surface epithelial cells, it is main to play
Protection and the effect for repairing epithelium.MUC16 is the important Tumor invasion for early diagnosing ovarian epithelial carcinoma, is widely used in facing
Bed.Recently research is found, MUC16 can promote oophoroma DISTANT METASTASES IN, the especially abdomen in oophoroma by being combined with mesothelin
Chamber shifts；With NK cells formation immunological synapse ovarian cancer cell can be helped to realize immunologic escape by suppressing ovarian cancer cell；
It can also convert, breed, migrate and shift to influence the Epithelial and stromal of tumour cell by the overexpression in ovarian cancer cell,
Promote the occurrence and development of oophoroma.Therefore, MUC16 can provide potential specific molecular target spot, or ovum for treatment of ovarian cancer
The immunization therapy of nest cancer provides new strategy.
Chimeric antigen receptor (chimeric antigen receptor, CAR) is a kind of antigen receptor of restructuring, simultaneously
With the function of combining antigen and immune cell activated.CAR T cell is transferred to, it, which is mainly characterized by obtaining, carries identification tumour
Antigen-specific receptor and the personalized curative effect for assigning T cell target killing activity.Therefore, using MUC16 as target spot chimeric antigen
Cellular immunotherapy new strategy based on acceptor can bring new hope to ovarian cancer patients.
At present, a series of CAR-T cells based on different tumor surface antigen designs show obvious anti-in vivo and in vitro
Tumor effect, especially target CD19 CAR-T cells achieved in the clinical patients of B cell malignant tumour it is good
Curative effect.But the T cell after these genetic modifications can show excessive activity, and it is difficult to control and may causes serious
Toxicity.
The content of the invention
In order to make up above deficiency, the invention provides a kind of the immune of the safety-type Chimeric antigen receptor modification of anti-MUC 16
Cell and its application.
The solution of the present invention is：
A kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, immunocyte are safety-type embedding through anti-MUC 16
Acceptor is closed to modify to obtain the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, the safety-type inosculating antibody of anti-MUC 16
Original receptor includes CD8leader, MUC16 land, CD8 hinge area, cross-film-stimulus structure domain, the conduction of CD3 ζ stimulus signals
Area and chemical induction element.
As preferable technical scheme, the cross-film-stimulus structure domain be selected from CD8, CD27, CD28, CD137/4-1BB,
All or part of fragment of CD134/OX40, ICOS molecule.
As preferable technical scheme, the chemical induction element is lured based on FKBP12-dimerization chemistry
Guide system (FK506binding protein 12-chemical inducers of dimerizations, FKBP12-
CID) build, include two copy FKBP12 mutant F36V-FKBP (Phe on 36 is substituted by Val), and in centre
Insert T2A sequences.
As preferable technical scheme, the chemical induction element is located at cross-film-stimulation in restructuring Chimeric antigen receptor
Between domain and CD3 ζ stimulus signal conducting regions.
As preferable technical scheme, the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 of the present invention is selected from
Autologous or transgenosis T cell, NK cells, cytotoxic T lymphocyte or regulatory T-cell, it is memory t cell, double special
Property T cell, CIK cell.
As preferable technical scheme, the fusion fragment of the safety-type Chimeric antigen receptor of anti-MUC 16 is
Leader-scFv (MUC16)-CD8-CD137-F36V-FKBP-T2A-F36V-FKBP-CD3 ζ fusion fragments.
As preferable technical scheme, Leader-scFv (the MUC16)-CD8-CD137-F36V-FKBP-T2A-
F36V-FKBP-CD3 ζ fusions fragment is the nucleotide sequence shown in sequence table SEQ ID NO.1.
The invention also discloses a kind of medicine for the treatment of cancer, the safety-type Chimeric antigen receptor modification containing anti-MUC 16 is exempted from
Epidemic disease cell.
The immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 is used for the use for preparing the medicine for the treatment of malignant tumour
On the way.
Preferable technical scheme, the malignant tumour are oophoroma.
By adopting the above-described technical solution, a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, exempts from
Epidemic disease cell is modified to obtain the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 through the safety-type Chimerical receptor of anti-MUC 16,
The safety-type Chimeric antigen receptor of anti-MUC 16 includes CD8leader, MUC16 land, CD8 hinge area, cross-film-stimulation
Domain, CD3 ζ stimulus signals conducting regions and chemical induction element.
The invention advantage：
1. the present invention provides the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, antigen spy can retained
The treatment function of small molecule regulation immunocyte is utilized on the basis of the opposite sex.Only in situation existing for heterodimeric small molecule
Under, the structure of the acceptor of division is able to completely so as to play a role.This drug regulation quantified enables the surgeon to accurately
Action intensity, time and the position of immunocyte are controlled, so as to mitigate toxicity.
2. the present invention is transformed in the second generation, third generation CAR technologies, chemical induction element is added, can protected
The toxic side effect for the treatment of is avoided while hindering therapeutic effect.And the present invention can be exempted from suitable for any by CAR technological transformations
Epidemic disease effector cell, greatly improve the Clinical efficacy and security of CAR technologies.
Brief description of the drawings
Fig. 1 is safety-type Chimeric antigen receptor principle design drawing of the present invention
Fig. 2 is Chimeric antigen receptor Leader-scFv (MUC16)-CD8-CD137-F36V-FKBP- of the present invention
The design drawing of T2A-F36V-FKBP-CD3 ζ fusion fragment.
Fig. 3 is slow virus Leader-scFv (MUC16)-CD8-CD137-F36V-FKBP-T2A- of the present invention
The schematic diagram of F36V-FKBP-CD3 ζ expression plasmids.
The efficiency that Fig. 4 is the expression CAR of CAR of the present invention (MUC16)-CIK cell is 37.9%.
Fig. 5 is CAR of the present invention (MUC16)-CIK in vitro to MUC16 positive ovarian cancerous cell lines OVCAR3 killing-efficiency
Increase with the increase of AP1903 concentration, the final concentration of 400nM killings rates of AP1903 reach more than 90%；As a result this is illustrated
CAR (the MUC16)-CIK cell for inventing design is acted on very High Fragmentation OVCAR3 cells, and activity is adjusted by AP1903
Control.
Fig. 6 is with the increase of AP1903 concentration in CAR of the present invention (MUC16)-CIK bodies to oophoroma tumor inhibiting rate
And increase, the injection without AP1903 after injection CAR (MUC16)-CIK cell, tumor mass becomes big, and growth tendency is not with injecting
CAR (MUC16)-CIK cell is completely the same.
Embodiment
In order that the technical means, the inventive features, the objects and the advantages of the present invention are easy to understand, tie below
Specific embodiment is closed, the present invention is expanded on further.
Embodiment 1 expresses the structure and virus of the slow virus plasmid of the Chimeric antigen receptor albumen of nucleotide coding of the present invention
Packaging
1. by fusion piece Leader-scFv (MUC16)-CD8-CD137-F36V-FKBP-T2A-F36V-FKBP-
CD3 ζ insertion Lentivirals pLent-C-GFP.
Leader-scFv (MUC16)-CD8-CD137-F36V-FKBP-T2A-F36V-FKBP-CD3 ζ CAR modules are shown
Opinion Fig. 2 (complete nucleic-acid sequences are shown in annex SEQ ID NO.1).
Leader-scFv (MUC16)-CD8-CD137-F36V-FKBP-T2A-F36V-FKBP-CD3 ζ each modules of CAR
Sequence
(1) sub- Leader nucleic acid artificial sequence (SEQ ID NO.2) is guided
(2) Anti-MUC16single chain Fv antibody (scFv) nucleic acid artificial sequence (SEQ ID NO.3)
(3) CD8Hinge areas nucleic acid artificial sequence (SEQ ID NO.4)
(4) CD8 transmembrane regions nucleic acid artificial sequence (SEQ ID NO.5)
(5) CD137 intracellular regions nucleic acid artificial sequence (SEQ ID NO.6)
(6) Linker nucleic acid artificial sequence (SEQ ID NO.7)
(7) F36V-FKBP nucleic acid artificial sequence (SEQ ID NO.8)
(8) autothermic cracking peptide T 2A nucleic acid artificial sequence (SEQ ID NO.9)
(9) CD3 ζ intracellular regions nucleic acid artificial sequence (SEQ ID NO.10)
Respectively by the nucleic acid artificial sequence, Anti-MUC16 nucleic acid artificial sequence, CD8Hinge areas for guiding sub- Leader
Nucleic acid artificial sequence, the nucleic acid artificial sequence of CD8 transmembrane regions, CD137 nucleic acid artificial sequence, Linker1 nucleic acid artificial sequence,
F36V-FKBP nucleic acid artificial sequence, autothermic cracking peptide T 2A nucleic acid artificial sequence, F36V-FKBP nucleic acid artificial sequence, Linker2
Nucleic acid artificial sequence, CD3 ζ nucleic acid artificial sequence entrust Sangon Biotech (Shanghai) Co., Ltd. to synthesize its whole table to reach
Frame, insert pLent-C-GFP carrier (Invitrogen) NotI-AsiSI sites (see Fig. 3), be transformed into E.coli (DH5 α), pass through
After sequencing is correct, simultaneously plasmid purification is extracted using the plasmid purification kit of Qiagen companies, obtains the height of recombinant expression carrier
Quality plasmid.
2. slow virus is packed, titre detection
Slow virus package cell line 293T is inoculated in containing in DMEM+10%FBS 10cm culture dishes, 37 DEG C, 5%
CO2Under the conditions of cultivate, adherent rate be 70%-80% after transfected.Recombinant plasmid and empty plasmid are packed with slow virus respectively
Plasmid uses calcium phosphate transfection method cotransfection 293T cells, specific method reference molecule clone.After transfection after 24h, cell is obvious
Increase, spherical in shape, nucleus becomes big, is rounded, and adherent ability declines and easy to fall off.Observed after 48h under inverted fluorescence microscope
To having egfp expression into the cell.After 72h, supernatant is collected, filtration sterilization, is saved backup in -80 DEG C of low temperature refrigerators.
Virus titer is determined according to Lenti-XTMGo StixTM kits (ocean Science and Technology Ltd. of Beijing China product), as a result table
It is bright, the titre 2.69 × 10 of recombinant slow virus6pfu/mL。
The slow-virus infection CIK cell of embodiment 2
The preparation of 1.CIK cells
75ml autologous patient peripheral bloods are taken, with TBD sample rates separating liquid (purchased from Tianjin Hao oceans China Tech biology), separation is outer
All blood mononuclear cells.(it is purchased from the culture medium of the recombinant interferon alpha-2a (being purchased from the pharmacy of the Shenyang three lives) containing 1000IU/ml
CORNING companies, 88-551-CM) for Fiber differentiation after 24 hours, the recombinant interleukin 2 for adding 1000IU/ml (is purchased from Shen
Positive three lives pharmacy), the induction of 50ng/ml OKT-3 and 5% autologous patient blood plasma continue culture 24 hours.Multiple proportions every three days
Liquid feeding, cultivate to the 14th day, the positive expression rate of CD3+, CD56+ in Flow cytometry CIK cell (CD3-FITC,
CD16/CD56-PE antibody is purchased from BECKMAN companies, A07735).CD3+ positive rates>The double positive rates of 80%, CD3+CD56+>
20%, it is considered as CIK and induces successfully, and leaves and takes the CIK and treat viral infection.
2. the amplification cultivation of CIK cell after slow-virus infection CIK cell and infection
CIK cell is infected with MOI=5 respectively with above-mentioned restructuring and unloaded slow virus.Metainfective 37 DEG C of cell, 5%CO2
After being cultivated 8 hours in incubator, cell is collected, rejoins virus liquid, 1000g, 32 DEG C, after centrifuging 90 minutes again, 37 DEG C,
5%CO2Continue to cultivate in incubator, multiple infection is so repeated, improve the efficiency of infection of CIK cell.2ml cultures are abandoned in suction
Supernatant, 2ml fresh CORNING culture mediums are added, continue to expand culture, culture is expanded to enough dosages to cell in 17 days.
(Fig. 4) is expressed by FC500 flow cytometers (being purchased from BECKMAN companies) FL1 Air conduct measurements Chimeric antigen receptor.To be uninfected by
CIK lymphocytes as negative control, its positive rate 37.9% of recombinant slow virus infection CIK cell.
The safety-type Chimeric antigen receptor of anti-MUC 16 is after being transferred to T cell and translating into albumen in the present invention, and T2A is by autotomying
Acceptor can be made to split into two parts, a part is antigen binding domain, and another part is intracellular signal transduction area.
Embodiment 3：CAR (MUC16) with chemical induction element-CIK cell killing activity research
1. the CAR (MUC16) with chemical induction element-CIK cell in vitro killing activity research
MUC16 positive ovarian cancerous cell line OVCAR3 as target cell, effector cell be CAR (MUC16)-CIK cell and
Unloaded slow-virus infection CIK.
It is inoculated with 100 μ l 1 × 105The target cell OVCAR3 in/hole is into 96 porocyte culture plates, with 1:2 effect targets are imitated than adding
Cell i.e. CAR-CIK cells are answered, and AP1903 is added according to final concentration of 0nM, 50nM, 100nM, 200nM, 400nM, effect is set
Answer cell controls group and target cell control group.It is placed in 5%CO2, 20 μ L CCK-8 are added per hole after 37 DEG C of incubator culture 24h,
Continue upper ELIASA after being incubated 2h to detect, the reading OD values at 450nm wavelength, killing rate=[(experimental group OD values-effect is thin by 1-
Born of the same parents control group OD values)/target cell control group OD values] × 100%.With unloaded slow-virus infection CIK, (result is shown in figure as a control group
5).CAR (MUC16)-CIK cell is to MUC16 positive ovarian cancerous cell lines OVCAR3 killing-efficiency with AP1903 concentration
Increase and increase, the final concentration of 400nM killings rates of AP1903 reach more than 90%；As a result the CAR that the explanation present invention designs
(MUC16)-CIK cell is acted on very High Fragmentation OVCAR3 cells, and activity is regulated and controled by AP1903.
2. killing activity research in CAR (MUC16)-CIK cell body with chemical induction element
18-22g female KM mices (being purchased from Traditional Chinese Medicine University Of Guangzhou) raise (23 ± 2 DEG C of room temperature, humidity 50% in Animal House
± 10%) the OVCAR3 cells of logarithmic phase, are collected, phosphate buffer (PBS) is diluted to 2 × 105Individual/mL.Under aseptic condition,
The left oxter inoculation 0.2mL OVCAR3 cell suspending liquids of mouse, observe 3-5d, treat that oxter the harder tubercle of grain of rice size occurs and made
To model successful standard.
(size that slide measure measures hypodermic tumour tissue block is 90-100mm to KM oophoroma Transplanted tumor models mouse3)
5 groups are randomly divided into, every group 20, starts to inject Experiment on therapy.Experimental group is respectively：
A. control group, the physiological saline of tail vein injections equal volume, continuous observation 14d；
B. one group is treated, tail vein injections 2 × 106An individual cell/CAR (MUC16)-CIK cell, 2 after injecting first
It carries out second and injected with dosage again, continuous observation 14d；
C. two groups are treated, tail vein injections 2 × 106An individual cell/CAR (MUC16)-CIK cell, 2 after injecting first
It carries out second and injected with dosage again, and locally injected into tumor 0.1mg/kg AP1903, continues after treatment injection 2h every time
Observe 14d；
D. three groups are treated, tail vein injections 2 × 106An individual cell/CAR (MUC16)-CIK cell, 2 after injecting first
It carries out second and injected with dosage again, and locally injected into tumor 0.5mg/kg AP1903, continues after treatment injection 2h every time
Observe 14d.
E. four groups are treated, tail vein injections 2 × 106An individual cell/CAR (MUC16)-CIK cell, 2 after injecting first
It carries out second and injected with dosage again, and every time after treatment injection 2h, locally injected into tumor 1mg/kg AP1903 are lasting to see
Examine 14d.
Survey body weight 1 time within every 3 days.Experiment is with 14 days for the cycle.After experiment terminates, dissection mouse separation tumour is weighed
(result is shown in Fig. 6).
And then injection CAR (MUC16)-CIK cell injects AP1903, the growth of mouse tumor is suppressed, and suppresses journey
Degree is directly proportional to AP1903 concentration.Injection without AP1903 after injection CAR (MUC16)-CIK cell, tumor mass becomes big, raw
Long trend is completely the same with not injecting CAR (MUC16)-CIK.
CAR (MUC16)-CIK cell that the conclusion present invention designs has obvious inhibitory action to KM mice tumors grews,
Action intensity, time and the position of CAR (MUC16)-CIK cell can be by AP1903 medicine accuracy controllings, so as to avoid CAR
(MUC16) toxic side effect of-CIK cell.
The safety-type Chimeric antigen receptor of anti-MUC 16 is under the sub- AP1903 effects of chemical induction in the present invention, F36V-FKBP bis-
Dimerization, the restructuring Chimeric antigen receptor of division reconfigures, and after complete acceptor is combined with specific antigen, can cause ITAM
Phosphorylation activation downstream albumen breeds T cell activation, to realize that "ON" acts on；Acted on without chemical induction, the restructuring of division
Chimeric antigen receptor can not breed T cell activation, to realize that "Off" acts on.
Embodiment 4：CAR (MUC16)-CIK cell clinical application method
For the treatment of MUC16 positive tumor patients, treatment method is local tumor injection and venous re-transfusion 2 × 106It is individual
CAR (MUC16)-CIK cell.Then local tumor injects AP1903 (0.01-100mg/kg) after treatment injection 2h.
General principle, principal character and the advantages of the present invention of the present invention has been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the simply explanation described in above-described embodiment and specification is originally
The principle of invention, without departing from the spirit and scope of the present invention, various changes and modifications of the present invention are possible, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent circle.
Sequence table
<110>Shandong Xing Rui bio tech ltd
<120>A kind of immunocyte of safety-type Chimeric antigen receptor modification of anti-MUC 16 and its application
<130> 2017
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agagtgaagt tcagcaggag cgcagacgcc cccgcgtaca agcagggcca gaaccagctc 60
tataacgagc tcaatctagg acgaagagag gagtacgatg ttttggacaa gagacgtggc 120
cgggaccctg agatgggggg aaagccgaga aggaagaacc ctcaggaagg cctgtacaat 180
gaactgcaga aagataagat ggcggaggcc tacagtgaga ttgggatgaa aggcgagcgc 240
cggaggggca aggggcacga tggcctttac cagggtctca gtacagccac caaggacacc 300
tacgacgccc ttcacatgca ggccctgccc cctcgc 336
Claims (10)
- A kind of 1. immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, it is characterised in that：Immunocyte is through anti- The safety-type Chimerical receptors of MUC16 are modified to obtain the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16, described anti- The safety-type Chimeric antigen receptors of MUC16 include CD8leader, MUC16 land, CD8 hinge area, cross-film-stimulus structure domain, CD3 ζ stimulus signals conducting regions and chemical induction element.
- 2. a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 as claimed in claim 1, its feature exist In：The cross-film-stimulus structure domain be selected from CD8, CD27, CD28, CD137/4-1BB, CD134/OX40, ICOS molecule wherein it One all or part of fragment.
- 3. a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 as claimed in claim 1, its feature exist In：The chemical induction element is built based on FKBP12-dimerization chemical induction subsystem, including two FKBP12 mutant F36V-FKBP is copied, and T2A sequences are inserted in centre.
- 4. a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 as claimed in claim 1, its feature exist In：The chemical induction element, which is located at the cross-film-stimulus structure domain of the Chimeric antigen receptor and the CD3 ζ, stimulates letter Between number conducting region.
- 5. a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 as claimed in claim 1, its feature exist In：The immunocyte is selected from autologous or transgenosis T cell, NK cells, cytotoxic T lymphocyte or regulatory T-cell, Memory t cell, bispecific T cell, CIK cell one kind therein.
- 6. a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 as claimed in claim 1, its feature exist In：The fusion fragment of the safety-type Chimeric antigen receptor of anti-MUC 16 is Leader-scFv (MUC16)-CD8-CD137- F36V-FKBP-T2A-F36V-FKBP-CD3 ζ fusion fragments.
- 7. a kind of immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 as claimed in claim 6, its feature exist In：Leader-scFv (the MUC16)-CD8-CD137-F36V-FKBP-T2A-F36V-FKBP-CD3 ζ fusion fragments For the nucleotide sequence shown in sequence table SEQ ID NO.1.
- A kind of 8. medicine for the treatment of cancer, it is characterised in that：Contain the safety-type inosculating antibody of anti-MUC 16 described in claim 1 The immunocyte of original receptor modification.
- It is pernicious that 9. the immunocyte of the safety-type Chimeric antigen receptor modification of anti-MUC 16 described in claim 1 is used for preparation treatment The purposes of the medicine of tumour.
- 10. the purposes of the medicine as claimed in claim 9 for preparing treatment malignant tumour, it is characterised in that：The malignant tumour For oophoroma etc..
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| CN110526991A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | Target Chimeric antigen receptor, the Chimeric antigen receptor T cell and its preparation method and application of FAP |
| CN110526977A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | It is a kind of to target the single-chain antibody of MUC1, Chimeric antigen receptor T cell and its preparation method and application |
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| CN110527667A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | A kind of Chimeric antigen receptor and Chimeric antigen receptor T cell and its preparation method and application targeting MUC16 |
| CN110526991A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | Target Chimeric antigen receptor, the Chimeric antigen receptor T cell and its preparation method and application of FAP |
| CN110526977A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | It is a kind of to target the single-chain antibody of MUC1, Chimeric antigen receptor T cell and its preparation method and application |
| CN110526988A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | A kind of Chimeric antigen receptor and Chimeric antigen receptor T cell and its preparation method and application targeting MUC1 |
| CN110526974A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | It is a kind of to target the single-chain antibody of MUC16, Chimeric antigen receptor T cell and its preparation method and application |
| CN110526979A (en) * | 2018-05-25 | 2019-12-03 | 深圳宾德生物技术有限公司 | Target single-chain antibody, the Chimeric antigen receptor T cell and its preparation method and application of FAP |
| JP2021525534A (en) * | 2018-06-04 | 2021-09-27 | プレシゲン，インコーポレイテッド | MUC16-specific chimeric antigen receptors and their use |
| WO2023217192A1 (en) * | 2022-05-10 | 2023-11-16 | 四川大学华西医院 | Preparation of chimeric antigen receptor immune cell constructed based on msln precursor protein and use thereof |
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