CN106117367A - A kind of HER 3 specific chimeric antigen receptor and application thereof - Google Patents
A kind of HER 3 specific chimeric antigen receptor and application thereof Download PDFInfo
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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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70596—Molecules with a "CD"-designation not provided for elsewhere
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- A—HUMAN NECESSITIES
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- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
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- A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
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- C07K14/7051—T-cell receptor (TcR)-CD3 complex
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2815—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD8
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
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- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/622—Single chain antibody (scFv)
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- C07—ORGANIC CHEMISTRY
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- C07K2319/00—Fusion polypeptide
- C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
- C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
Abstract
The invention discloses a kind of HER 3 specific chimeric antigen receptor, described Chimeric antigen receptor is made up of people's anti-HER 3 single-chain antibody, the intracellular signal structures in series of people antibody CD8TM, CD137, and CD3 ζ.The invention also discloses aminoacid sequence and the nucleotide sequence of above-mentioned HER 3 specific chimeric antigen receptor.This Chimeric antigen receptor is used for modifying T lymphocyte, and the lymphocyte after modification can be used for the treatment of HER 3 gene expression related neoplasms.
Description
Technical field
The present invention relates to cellular immunization technical field, particularly relate to a kind of HER-3 specific chimeric antigen receptor and answer
With.
Background technology
The adoptive cell therapy of tumor (Adoptive cell therapy, ACT) is by self or the antitumor effect of allosome
Answering the precursor of cell, use IL-2, anti-CD49d McAb in vitro, the activator such as specific polypeptide carries out inducing, activate and expanding
Increase, then transfer to tumor patient, improve patient's antineoplastic immune power, to reach to treat and prevent the purpose of recurrence.Adoptive
Immune effector cell treatment is subject to people's attention, for the most alive in nearly more than ten years immunotherapy of tumors because of the advantage of tool uniqueness
The research field jumped, achieves, in antineoplastic is treated, the achievement attracted people's attention.ACT goes through NK, gamma delta T, CD3AK, DC-
The developmental stage such as CIK, LAK, TIL, CIK, EAAL, its curative effect, specificity, are integrated with the feelings of the aspect such as efficiency, side effect reaction
Condition is progressively improved.And the activation of T cell needs the stimulation of two signals.The TCR-CD3 complex on T cell surface and antigenic peptides-
MHC combines, it is provided that the first signal of T cell activation；The costimulatory molecules on T cell surface combines with corresponding part, it is provided that T
The secondary signal of cell activation.Therefore reply T cell is transformed.Use genetic modification method to prepared the most in vitro
Cell carry out φt cell receptor (T cell receptor, TCR) and CAR genetic modification.Although being both the main flow side of tumor ACI
To, but Chimeric antigen receptor (chimeric angiten receptor, CAR) is modified T cell and is modified T cell with TCR
(TCR-T) still making a big difference between therapy, Pros and Cons is different, and wherein the sharpest edges of CAR-T are it
The performance of antitumor action does not relies on the selection of HLA molecule and tumor antigen target and is not limited to protein substance.
In recent years, swelling based on Chimeric antigen receptor (chimeric angiten receptor, CAR) modifies T cell
Tumor adoptive immunotherapy makes substantial progress.T cell after modification not only has a killing ability of targets neoplastic cells, and can gram
Take tumor by local immunosuppressant microenvironment and break the state of host immune tolerance.CAR, by continuously improving, has sent out
Forth generation has been arrived in exhibition.First generation CAR is mainly made up of scFv and ITAM (usually CD3 ζ) identifying tumor associated antigen, but
Due to only CD3 ζ activation signals, T cell can not fully be rised in value, and the T cell time-to-live in vivo of activation is short.The second filial generation
CAR adds costimulatory molecules such as CD28, CD134, a CD137 etc. on the basis of generation CAR, enhance t cell activation,
Propagation, internal hold time, thus improve antineoplastic effect.Third generation CAR be re-introduced on the basis of secondary CAR one with
On costimulatory molecules.Forth generation CAR (TRUCKS, T cells redirected for universal cytokine
Killing) mainly by cytokines such as introducing IL-12, IL-23, IL-27, inherent immunity cell-macrophage etc. is called together to kill
Hinder the negative tumor cell of TAA.
According to the difference of T cell position residing for CAR, CAR is broadly divided into three parts: born of the same parents' exoantigen land, cross-film district, born of the same parents
Interior signal transduction district.Wherein born of the same parents' exoantigen land is typically by for tumor associated antigen (tumor associated
Antigen, TAA) the single-chain variable sequence (single chain fragment variable, scFv) of monoclonal antibody
Constitute (light and variable region of heavy chain connects formation by hinge region).This partial function is to identify tumor associated antigen, and CAR-T is thin
Born of the same parents are attached to tumor up.Cross-film district is generally by dimer protein compositions such as CD3, CD28, CD8.Intracellular signal transduction district is main
For ITAM (immunoreceptor tyrosine-based activation motifs,
ITAM, usually CD3 ζ or Fc ε RI γ).Its major function is to T cell signal transduction, improves antitumor and cytokine
Secretion capacity.
CAR-T cellular immunotherapy is distinguished as the operation in traditional tumour treatment, radiotherapy, and chemotherapy shows huge excellent
Gesture.Being embodied in killing tumor accuracy high, CAR-T cell therapy uses the technology that antigen and antibody specific combines, only
The tumor cell that TSA is expressed is killed, little to Normocellular injury；Killing range of tumor is extensive, as long as expressing
Tumor associated antigen, CAR-T cell just can be removed, the most effective for metastatic tumo(u)r recurrent tumor；Keep away for patients
Exempt from the misery of chemicotherapy, got well rapidly.
HER-3 (human epidermal growth factor receptor3) is also referred to as ErbB3, is I type cross-film cheese ammonia
Acid kinase receptor, is tyrosine kinase receptor ErbB one unique member of family.Its function, regulation activity and cancer target
The sensitivity for the treatment of is closely bound up with the dimerization of other members of ErbB family.Several Kinds of Malignancy such as breast carcinoma, Colon and rectum
The common squamous cell carcinoma of the head and neck (scale cancer) of cancer, uveal, stomach, ovary, prostate, bladder cancer all height are expressed
HER-3.Because it in cancerous tissue and the wide expression of critical function in cancer progression, is attempted Targeted cancer therapy ErbB family and becomes
Member has become widely studied emphasis.The preparation CAR-T with HER-3 as target spot has highly important meaning for treatment tumor
Justice.
Summary of the invention
The technical problem existed based on background technology, it is an object of the invention to provide a kind of HER-3 specific chimeric and resists
Original receptor.
The present invention also aims to provide aminoacid sequence and the nucleotide sequence of above-mentioned Chimeric antigen receptor.
The present invention also aims to provide the application of above-mentioned Chimeric antigen receptor.
To achieve these goals, a kind of HER-3 specific chimeric antigen receptor that the present invention proposes, by the anti-HER-3 of people
Single-chain antibody, the intracellular signal structures in series of people antibody CD8TM, CD137, and CD3 ζ is constituted.
Preferably, the aminoacid sequence of described Chimeric antigen receptor is as shown in SEQ ID NO.1.
Preferably, the nucleotide sequence of described Chimeric antigen receptor is as shown in SEQ ID NO.3.
Preferably, the aminoacid sequence of described people anti-HER-3 single-chain antibody is as shown in SEQ ID NO.4.
Preferably, the nucleotide sequence of described people anti-HER-3 single-chain antibody is as shown in SEQ ID NO.5.
Preferably, described people anti-HER-3 single-chain antibody includes variable region of light chain and variable region of heavy chain, described variable region of light chain
Nucleotide sequence as shown in SEQ ID NO.2.
Preferably, the nucleotide sequence of described variable region of heavy chain is as shown in SEQ ID NO.10.
Preferably, there are a connection peptides, its ammonia between heavy chain molecule and the light chain molecule of described people anti-HER-3 single-chain antibody
Base acid sequence is Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser.
Preferably, the amino terminal of described Chimeric antigen receptor contains a signal peptide, the aminoacid sequence of described signal peptide
Row are as shown in SEQ ID NO.6.
Preferably, the aminoacid sequence of CD137 is as shown in SEQ ID NO.7.
Preferably, the nucleotide sequence of CD137 is as shown in SEQ ID NO.8.
Preferably, the domain that the intracellular signal structures in series of people's antibody CD8TM, CD137, and CD3 ζ is constituted is T cell
Costimulatory signal conducting structure, its aminoacid sequence is as shown in SEQ ID NO.9.
The above-mentioned HER-3 specific chimeric antigen receptor that the present invention also proposes is at preparation treatment HER-3 related neoplasms medicine
In application.
Wherein, relevant for HER-3 tumor includes breast carcinoma, colon and rectum carcinoma, gastric cancer, ovarian cancer, carcinoma of prostate, wing
Guang cancer, uveal etc..
The present invention filters out HER-3 monoclonal antibody, codon optimization, gene according to laboratory from human antibody storehouse
Synthesize the ScFv sequence of anti-HER-3, and synthesis ScFv is cloned in pSFG-CD8TM-CD137-CD3 ζ, be i.e. built into HER-
3-scFv-CD8TM-CD137-CD3 ζ Chimeric antigen receptor, i.e. obtains Chimeric antigen receptor HER-3-scFv-of the present invention
CD8TM-CD137-CD3ζ。
Chimeric antigen receptor and slow virus packaging plasmid PeqPam3 and RD114env is utilized to be packaged into slow disease at 293T cell
Poison, utilizes this slow virus infection T lymphocyte.Utilize Elisa detection CAR-T cell IFN-γ under antigenic stimulus secretion and
The lethal effect of the tumor cell that CCK8 method is positive to HER-3.
The invention discloses the structure of the specific Chimeric antigen receptor of HER-3, by Infection in Vitro, modify T lymph thin
Born of the same parents, the T lymphocyte after modification discharges the killing tumor cells such as IFN-γ.The specific Chimeric antigen receptor of HER-3 can be used
Targeted therapy in HER-3 related neoplasms.
Accompanying drawing explanation
Fig. 1 is the electrophoretogram of the fragment of the embodiment of the present invention 1 gained HER-3scFv mesh, and wherein M is normalization markers.
Fig. 2 is the electrophoretogram of the fragment of the embodiment of the present invention 1 gained CD8TM-CD137-CD3 ζ mesh, and wherein M is standard mark
Note thing.
Fig. 3 is to use Western Blotting to detect gained HER-3 specific chimeric of the present invention in the embodiment of the present invention 2
Expression after antigen receptor transfection 293T cell.
Fig. 4 is the expression of HER-3 in the exponential phase tumor cell that Western Blotting detection is different.
Fig. 5 is that the killing of tumor cell is examined by the T cell that gained HER-3 specific chimeric antigen receptor of the present invention is modified
Survey.
Fig. 6 is that the T cell of gained HER-3 specific chimeric antigen receptor of the present invention modification is by HER-3 antigenic stimulus
The expression of rear IFN-γ.
Detailed description of the invention
Below, by specific embodiment, technical scheme is described in detail.
The structure of embodiment 1:HER-3 specific chimeric antigen receptor slow virus carrier
1. utilize the Fab sequence of the anti-HER-3 of this laboratory screening, select the sequence of wherein VH and VL, be used for designing CAR
ScFv sequence in carrier, uses Jin Sirui biotech company OptimumGeneTM gene design software to carry out codon
Optimize further, at the sequence two ends optimized respectively plus NcoI, BamHI restriction enzyme site, the sequence after optimization by Nanjing gold this
Auspicious biotech company completes gene chemical synthesis, and the sequence after synthesis is cloned in pUC57 carrier, the named HER-3-scFv-of plasmid
pUC57。
2.HER-3-scFv-pUC57 with restricted enzyme NcoI and BamH I enzyme action, enzyme action system is as follows: 2 μ g
HER-3-scFv-pUC57,1 μ L NcoI, 1 μ L BamH I, 2 μ L 10 × enzyme cutting buffering liquids, mend 20 μ L, 37 DEG C of water-baths with water
In overnight, digestion products, with the agarose gel electrophoresis of 1%, cuts purpose band under ultraviolet, uses DNA gel to reclaim reagent
Box (AXYGEN company) reclaims purpose fragment；Its result is as it is shown in figure 1, pUC57-HER-3-ScFv carrier is through NcoI and BamH I
Enzyme action release purpose band.
With same method NcoI and BamH I enzyme action pSFG-CD8TM-CD137-CD3 ζ carrier, with agarose gel electricity
Swimming, reclaims purpose fragment.Its result is as in figure 2 it is shown, pSFG-CD8TM-CD137-CD3 ζ carrier is released through NcoI and BamH I enzyme action
Put purpose band.
HER-3-ScFv after reclaiming is connected by T4DNA ligase with enzyme action carrier, and reaction system is as follows: 2 μ L
HER-3-ScFv, 2 μ L carrier digestion products, 1 μ L 10 × connection buffer, 1 μ L ligase, mend 10 μ L, 16 DEG C of water-baths with water
In overnight.Take connection product to join in DH5TM competence and [to prepare with reference to E. coli competent in " molecular cloning " third edition
(CaCl2Method)], place incubated overnight in 37 DEG C of bacteriological incubators.The single bacterium colony of picking, amplification culture, extract positive colony
Plasmid, through enzyme action and order-checking, by correct carrier named HER-3-scFv-CD8TM-CD137-CD3 ζ.
Embodiment 2:HER-3 specific chimeric antigen receptor expression identification
Retroviral vector is extracted with reference to without the operating instruction of (sky root is biological) in the big extraction reagent kit of endotoxin plasmid
HER-3-scFv-CD8TM-CD137-CD3 ζ, the plasmid of extraction, it is transfected in HEKC 293T with PI transfection reagent, 48h
After, wash one time with PBS, extract reagent (RIPA) cell lysis with cell protein, extract the albumen warp of the 293T cell after transfection
After the SDS-PAGE of 10% separates, constant current (300mA, 1h) is transferred on pvdf membrane, incubates with AntiCD3 McAb ζ (1:1000) antibody
Educate, 4 DEG C of overnight incubation.After washing 3 times with PBST, by the two of HRP sheep anti-Mouse anti-(1:5000) incubated at room 1h.Add ECL
After colour developing, carrying out imaging with the ChemiDoc XRS System of Bio-Rad company, its result is as shown in Figure 3.
As shown in Figure 3: the recombiant plasmid constructed by the present invention is able to detect that the expression of purpose band, size and sun ginseng
CD19 is consistent, and the 293T cell of untransfected does not has band.
The preparation of the T lymphocyte that embodiment 3:HER-3 specific chimeric antigen receptor is modified
1. the packaging containing anti-HER-3 Chimeric antigen receptor slow virus
Load is packed with extracting retrovirus without the operating instruction of (sky root is biological) in the big extraction reagent kit of endotoxin plasmid
Body RD114 and HER-3-scFv-CD8TM-CD137-CD3 ζ retroviral vector cotransfection in GP2-293T cell, transfection
Rear 48h collects cell conditioned medium, and 4000rpm is centrifuged 10min, removes the cell in supernatant and cell debris.Filter membrane by 0.45 μm
Filtering, subpackage is frozen, standby.
The preparation of 2.T lymphocyte
Take the fresh anticoagulation of 20mL healthy volunteer, by lymph separation liquid (GE company) discrete peripheral blood lymphocytes
(PBMC).Cell CD3 and CD28 flat board after separation stimulate 48h, and with T lymphocytes culture medium GT-T551, (TAKARA is public
Department) add 3% self blood plasma and induce, obtain T lymphocyte.
The preparation of 3.CAR-T cell
Being coated non-tissue culturing plate 24 orifice plate with the RetroNectin (TAKARA company) of 50 μ g/mL, every hole adds 500 μ
L, 4 DEG C overnight.Every hole adds 500 μ L viral supernatants, hatches 30min for 37 DEG C.Remove viral supernatants, add 500 μ L viruses
Supernatant, hatches 30min for 37 DEG C, and except viral supernatants, every hole adds 1.5mL viral supernatants, adds the T lymph after 0.5mL dilution
Cell.Thus obtain HER-3-scFv-CD8TM-CD137-CD3 ζ T cell i.e. HER-3 specific C AR-T cell.
The embodiment 4:HER-3 specific C AR-T cell lethal effect to HER-3 related neoplasms
The expression of HER-3 in 1.Western Blotting detection tumor cell
Choose the human melanoma cell A375 of exponential phase, human breast cancer cell MDA-MB-231 and human breast carcinoma thin
Born of the same parents MCF-7, extracts reagent (RIPA) cell lysis with cell protein, extracts cell protein, carries out Western Blotting inspection
Survey, testing result as shown in Figure 4: human breast cancer cell MDA-MB-231 and human breast cancer cell line Bcap-37 detect the table of HER-3
Reach, and human melanoma cell A375 can't detect the expression of HER-3.
The lethality detection of 2.CAR-T cells against tumor
Tumor cell culture medium is adjusted to 5 × 106/ mL, every hole 50 μ L, by E:T (effector lymphocyte and target cell ratio)
For 16:1,8:1,4:1,2:1, it is separately added into tumor cell 2.5 × 106Individual, 1.25 × 106Individual, 6.25 × 105Individual, 3.125 ×
105Individual；After cell is the most adherent, collecting T cell and CAR-T cell, adjusting cell concentration is 1 × 106/ mL, every hole 50 μ L,
Cultivate 12h.Abandon supernatant and add the CCK8 of 100 μ L dilutions, hatch 4～6 hours, the light absorption value of microplate reader detection OD450.
Above-mentioned tumor cell is human melanoma cell A375, human breast cancer cell MDA-MB-231 and human breast cancer cell
MCF-7。
Testing result is as shown in Figure 5: CAR-T cell to the killing rate of the tumor cell that HER-3 expresses higher than HER-3 not table
The tumor cell reached；CAR-T cells against tumor cells lethal effect is higher than T cell, higher than media alone.
3.ELISPOT detection CAR-T cell IFN-γ after by HER-3 antigenic stimulus is expressed
With aseptic be coated liquid dilution IFN-γ antibody after add ELISPOT (Millipore, Cat.No.MAIPS4510) put down
In plate, every hole 100 μ L, 4 DEG C stand overnight.Second day with the aseptic liquid that is coated by Plate wash 2 times.It is complete that every hole adds 200 μ L
Full culture medium, room temperature closes 1h.Wash away the cell culture fluid RPMI-1640 containing serum, wash three times with PBS.Prepare outside HER-3 born of the same parents
District's polypeptide, is diluted in the cell culture fluid RPMI-1640 containing serum, is adjusted to final concentration 1 μ g/mL, every hole 100 μ L.Adjust to be checked
Surveying CAR-T cell concentration is 1 × 106/ mL, every hole adds 100 μ L, 37 DEG C, 5%CO2Place 24h, discard cell and culture medium,
Wash 3 times with ELISPOT cleaning mixture.Every hole adds biotin labeled detection antibody, and 100 μ L/ holes, 4 DEG C stand overnight.Use again
ELISPOT cleaning mixture washs 4 times.Every hole adds the Avidin of HRP labelling, 100 μ L/ holes, and room temperature places 45min.Use ELISPOT
After cleaning mixture washs 3 times, wash 2 times with PBS.Every hole adds 100 μ L ACE chromogenic substrates, color development at room temperature 20～60min.Treat out
The most substantially after collection drop point, wash 3 times with sterilized water and terminate reaction.Air dries flat board, calculates with ELISPOT plate reading machine and formed
Speckle number, its result is as shown in Figure 6.As shown in Figure 6: CAR-T cell can be secreted under specificity HER-3 antigenic stimulus
IFN-γ。
The above, the only present invention preferably detailed description of the invention, but protection scope of the present invention is not limited thereto,
Any those familiar with the art in the technical scope that the invention discloses, according to technical scheme and
Inventive concept equivalent or change in addition, all should contain within protection scope of the present invention.
Claims (10)
1. a HER-3 specific chimeric antigen receptor, it is characterised in that described Chimeric antigen receptor is by people's anti-HER-3 strand
Antibody, the intracellular signal structures in series of people antibody CD8TM, CD137, and CD3 ζ is constituted.
HER-3 specific chimeric antigen receptor the most according to claim 1, it is characterised in that described Chimeric antigen receptor
Aminoacid sequence is as shown in SEQ ID NO.1.
HER-3 specific chimeric antigen receptor the most according to claim 1 or claim 2, it is characterised in that described Chimeric antigen receptor
Nucleotide sequence as shown in SEQ ID NO.3.
4. according to HER-3 specific chimeric antigen receptor described in any one of claim 1-3, it is characterised in that described people resists
The aminoacid sequence of HER-3 single-chain antibody is as shown in SEQ ID NO.4；Preferably, the nucleic acid of described people anti-HER-3 single-chain antibody
Sequence is as shown in SEQ ID NO.5.
5. according to HER-3 specific chimeric antigen receptor described in any one of claim 1-4, it is characterised in that described people resists
HER-3 single-chain antibody includes variable region of light chain and variable region of heavy chain, the nucleotide sequence of described variable region of light chain such as SEQ ID NO.2
Shown in；Preferably, the nucleotide sequence of described variable region of heavy chain is as shown in SEQ ID NO.10.
6. according to HER-3 specific chimeric antigen receptor described in any one of claim 1-5, it is characterised in that described people resists
Having a connection peptides between heavy chain molecule and the light chain molecule of HER-3 single-chain antibody, its aminoacid sequence is Gly-Gly-Gly-
Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser。
7. according to HER-3 specific chimeric antigen receptor described in any one of claim 1-6, it is characterised in that described inosculating antibody
The amino terminal of original receptor contains a signal peptide, and the aminoacid sequence of described signal peptide is as shown in SEQ ID NO.6.
8. according to HER-3 specific chimeric antigen receptor described in any one of claim 1-7, it is characterised in that the amino of CD137
Acid sequence is as shown in SEQ ID NO.7；Preferably, the nucleotide sequence of CD137 is as shown in SEQ ID NO.8.
9. according to HER-3 specific chimeric antigen receptor described in any one of claim 1-8, it is characterised in that people's antibody
The domain that the intracellular signal structures in series of CD8TM, CD137, and CD3 ζ is constituted is T cell costimulatory signal conducting structure, its
Aminoacid sequence is as shown in SEQ ID NO.9.
10. as described in any one of claim 1-9, HER-3 specific chimeric antigen receptor treats HER-3 related neoplasms in preparation
Application in medicine.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110526990A (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 CD30 |
| 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 |
| 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 |
| CN110526986A (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 CD30 |
| 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 |
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| WO2014160627A1 (en) * | 2013-03-25 | 2014-10-02 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Anti-cd276 polypeptides, proteins, and chimeric antigen receptors |
| CN105330750A (en) * | 2015-11-20 | 2016-02-17 | 上海细胞治疗研究院 | Molecular brake for rapidly stopping killing effect of CAR-T (T cell engineered with chimeric antigen receptors) and application of molecular brake |
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