CN103214556B - Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1) and application thereof - Google Patents
Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1) and application thereof Download PDFInfo
- Publication number
- CN103214556B CN103214556B CN201310112763.0A CN201310112763A CN103214556B CN 103214556 B CN103214556 B CN 103214556B CN 201310112763 A CN201310112763 A CN 201310112763A CN 103214556 B CN103214556 B CN 103214556B
- Authority
- CN
- China
- Prior art keywords
- peptide
- leu
- epitope peptide
- cell
- antigen
- 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
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention discloses an anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1). The epitope peptide comprises a nonapeptide, the amino acid sequence of which is P22: Tyr-Leu-Ile-Arg-Arg-Ile-Glu-Glue-Leu. The invention further comprises an application of the peptide in preparing tumor treating polypeptide vaccine. The invention has the advantages that the epitope peptide of metastasis associated antigen is initially evaluated by a theory and experiment combined method. Evaluated nonapeptide is not reported in documents, so that the invention provides a theoretical base for developing tumor treating polypeptide vaccines based on antigen MTA1 and lays a foundation for construction of subsequent multivalent antigen peptide vaccines, long peptide vaccines, multi-epitope vaccines and the like.
Description
Technical field
The present invention is that patent application is number of patent application: 201110138635.4, and the applying date: on 05 26th, 2011, denomination of invention was the divisional application of " antitumor CTL epitope peptide and the application thereof in MTA1 source ".
Technical field
The present invention relates to peptide, especially relate to the antitumor CTL epitope peptide in MTA1 source, the invention still further relates to this peptide in the application of preparing in tumor therapeutic polypeptide vaccine.
Background technology
Transfer is a complex process that relates to many reasons and product thereof, comprises that tumour cell departs from from primary tumor, invades lymphatic vessel, blood vessel and surrounding tissue, and induction of vascular forms, and escapes the antitumor reaction of host cell and grows at metastasis site.Therefore, understanding the related gene of metastases and gene product is the heat topic of studying both at home and abroad at present.
Metastasis related protein 1(MTA1) be that the people such as Toh in 1994 identify discovery in people's breast cancer cell.It crosses research discovery in recent years and expresses in multiple metastatic tumo(u)r, as mammary cancer, gastrointestinal cancer, the esophageal carcinoma, carcinoma of the pancreas, cancer of the stomach, liver cancer, nonsmall-cell lung cancer etc., and low expression or do not express in healthy tissues, and its expression level and its transfer in tumor tissues or infiltrate potential and have obvious relation.These results of study show that MTA1 is an extraordinary tumor associated antigen, are the target spots of tumor diagnosis and therapy.Sight is concentrated on the especially aspect such as expression, molecular function of MTA1 in multiple mankind tumor tissue and tumour cell of MTA family member by most researchers in recent years, and the people such as Li Kai are obtaining certain achievement in research aspect the anti-prostate cancer vasculogenesis of MTA1.Have not yet to see the report for the HLA-I class restricted CTL epitope peptide of MTA1.
Along with immunologic development of modern times, the vital role that cytotoxic T lymphocyte (Cytotoxic T lymphocyte, CTL) is brought into play in tumour more and more comes into one's own.How effectively to excite the specificity cellular immunity response of CTL mediation, bring into play antitumor usefulness, become an important topic in current tumor therapeutic polypeptide vaccine development field.Previously research is found, causes CTL immune response, and incomplete tumour antigen molecule, but the specific CTL epi-position (Epitope) in antigen source.After antigen presenting cell picked-up tumour antigen, being processed into by proteolyzing is the polypeptide fragment of 8～10 amino acid lengths, it is CTL epi-position, and then major histocompatibility complex class I (MHC-I) quasi-molecule in endoplasmic is combined and is formed polypeptide-MHC mixture (peptide-MHC complex, pMHC), and the most at last pMHC submission to cell surface φt cell receptor (the T cell receptor for CD8+T cell surface, TCR) identification, thereby activation CTL cell, causes CTL immunne response.
Summary of the invention
The object of the present invention is to provide a kind of antitumor CTL epitope peptide tumour cell SW620 to the MTA1 source of certain lethality, the present invention simultaneously also provides this peptide in the application of preparing in tumor therapeutic polypeptide vaccine.
For achieving the above object, the present invention can take following technical proposals:
The antitumor CTL epitope peptide in MTA1 of the present invention source, is made up of nine amino-acid residues, and the aminoacid sequence of described epitope peptide is
P22:Tyr-Leu-Ile-Arg-Arg-Ile-Glu-Glu-Leu molecular weight is 1204.49.
The present invention also comprises that above-mentioned peptide preparing the application of tumor therapeutic polypeptide vaccine of this antigen of high expression level.
The invention has the advantages that and adopt method preliminary evaluation theoretical and that experiment combines to go out the epitope peptide of metastases related antigen.The nonapeptide of identifying has no bibliographical information, for the tumor therapeutic polypeptide vaccine of development based on antigen MTA1 provided fundamental basis, and lays the foundation for the structure of the antigen peptide vaccine of follow-up multivalence, long peptide vaccine, polyepitope vaccines etc.
Accompanying drawing explanation
Fig. 1-5th, the mass spectroscopy figure of epitope peptide of the present invention.
Fig. 6 is the specific CTL of the epitope peptide of the present invention induction lethal effect to tumour cell SW620 (the MTA1-positive, the HLA-A2 positive).
Fig. 7-11st, the ability of the specific CTL secretion of gamma-IFN of epitope peptide induction of the present invention.
Embodiment
The antitumor CTL epitope peptide in MTA1 of the present invention source, comprises nonapeptide, and the aminoacid sequence of described nonapeptide is
P22：Tyr-Leu-Ile-Arg-Arg-Ile-Glu-Glu-Leu
Or P57:Ala-Leu-Ala-Asp-Lys-His-Ala-Thr-Leu
Or P109:Phe-Leu-Ser-Arg-Gln-Leu-Glu-Ser-Leu
Or P129:Thr-Leu-Leu-Asn-Glu-Thr-Glu-Ser-Leu
Or P173:Tyr-Gln-Ala-Asp-Ile-Thr-Asp-Leu-Leu.
The present invention mainly adopts the method for integrating theory with practice, according to the primary structure of antigen, adopt Immunoinformatics means, use SYFPEITHI, BIMAS, NetCTL 1.2 and IEDB database to carry out forecast analysis to the HLA-A*0201 restricted CTL epitope of MTA1 proteantigen.
Screening obtains above-mentioned epitope peptide and adopts the Fmoc scheme of standard to synthesize, and after HPLC purifying, its purity is greater than 90%, and mass spectroscopy also confirms that its molecular weight meets theoretical value.Each peptide Mass Spectrometric Identification figure is shown in Fig. 1-5.
Synthetic and the preparation of above-mentioned epitope peptide: adopt solid-phase synthesis.Basic procedure is as follows: first an amino is connected on insoluble solid phase carrier Wang resin by the amino acid of Fmoc radical protection, and the then protecting group of desamidizate, first amino acid be connected on solid phase carrier; Secondly amino is activated with condensing agent by second amino acid whose carboxyl of Fmoc radical protection; amino acid after activation reacts with first the amino acid whose amino that is connected on solid phase carrier and forms peptide bond, now on solid phase carrier, has just generated a dipeptides with protecting group.Repeat above-mentioned peptide bond and form reaction, make peptide chain from C end to the growth of N end, until reach needed peptide chain length, finally cutting obtains object peptide.After HPLC purifying, its purity is greater than 90%, and mass spectroscopy also confirms that its molecular weight meets theoretical value.(referring to: 1. only moral, Chen Changqing work of Huang, polypeptide is synthetic, Science Press, 1985.2. .N. Xiu Ede, H.D. Jia Kubuke work, Liu Keliang etc. translate, peptide: chemistry and biology, Science Press, 2005).
The separation of the human peripheral blood single nucleus cell (PBMCs) of epitope peptide of the present invention detects with ELISPOT experiment: the peripheral blood that extracts health donors separates through density gradient centrifugation, obtains PBMCs, adds interleukin II (IL-2, Peprotech company) and people β
2microglobulin induction differentiation CTL cell, further LDH and ELISPOT verify in vitro.Method is as follows:
1, the separation of PBMCs and induction: (1) is with 40ml PBS(PH 7.2) dilution anti-freezing peripheral blood 40ml after treatment; (2) in centrifuge tube, add 4ml Lymphoprep parting liquid (Axis-Shield company); (3) add in 8ml step (1) peripheral blood after dilution on 4ml Lymphoprep parting liquid liquid level; (4) 20 ℃ centrifugal (2000rmp × 20min); (5) after centrifugal, be divided into four layers, discard the superiors, drawing the second layer with glass pipette is tunica albuginea layer (being rich in PBMCs); (6) PBS(pH 7.2 for tunica albuginea layer of sucking-off) twice of centrifuge washing; (7) with 24 orifice plate bed boards, the concentration of cell is 1 × 10
6/ ml, every hole 1ml; Within (8) second days, every hole adds 3 μ g people β
2microglobulin and 10 μ g epitope peptides set up PBS group (substituting epitope peptide using PBS) as negative control group simultaneously; Within (9) the 3rd days, every hole adds 50u IL-2; Within every 2～3 days, change liquid, after Yu Qitian, carrying out the second wheel load peptide (is that 50u IL-2,10 μ g people β are added in every hole
2microglobulin and 10 μ g epitope peptide/PBS), after fortnight, carry out third round lotus peptide.After third round lotus peptide 3 days, obtain effector cell CTL, then carry out the effect of LDH experiment and ELISPOT experiment detection peptide to be measured.
2, LDH detects: use CytoTox 96 Non-Radioactive Cytotoxicity Assay(cytotoxicity detection kit, Promega company) carry out LDH test and detect cytotoxic activity.Step following (referring to test kit specification sheets):
1) set up check-out console (100 μ l/ hole)
(1) set up experimental group: using tumour cell EC-9706(ATCC company) as target cell (optimum target cell number: 5000) add above-mentioned effector cell CTL by difference effect target than 10:1,20:1,40:1;
(2) set up the spontaneous release group of effector cell;
(3) set up the spontaneous release group of target cell;
(4) set up the maximum release group of target cell;
(5) set up volume correction control group;
(6) set up background control group.
2) lysis and results supernatant
(1) 37 ℃, 5%CO
2hatch check-out console (5h);
(2) in the maximum release group of target cell and volume correction control group, add lysate, in every 100 μ l substratum, add 10 μ l lysates (10 ×), before results supernatant, 45min adds lysate;
(3) the centrifugal 4min of 250g, results supernatant.
3) LDH detects
(1) transferase 45 0 μ l supernatant is to another orifice plate;
The substrate mixed solution of dilution is added rapidly in (2) 50 μ l/ holes, and room temperature lucifuge is hatched 30min;
(3) add 50 μ l stop baths;
(4), by the bubble removal containing in hole, detect 490nm absorption value OD in one hour.
Cell killing rate calculation formula is as follows:
Kill rate (%)=［ (OD
experimental group-OD
the spontaneous release group of effector cell-OD
the spontaneous release group of target cell)/(OD
the maximum release group of target cell-OD
the spontaneous release group of target cell) × 100%
(note: the absorption value of all experimental group, the spontaneous release group of effector cell, the spontaneous release group of target cell all should subtracting background mean absorbance; The maximum release group of target cell absorption value should deduct the mean absorbance of volume correction control group)
The results are shown in Figure 6, as shown in Figure 6, the specific CTL of epitope peptide induction is to the tumour cell SW620(HLA-A*0201-positive, the MTA1-positive) all demonstrate certain kill rate.
3, ELISPOT experiment: specific experiment step is as follows: (1) sealing: take out required lath, with the substratum sealing containing 5% FCS RPMI 1640 substratum, 200 μ L/holes, under room temperature, (25 ℃) are deducted (FCS can seal the FCS acceptor of institute's coated antibody to reduce nonspecific reaction) after leaving standstill 5 ~ 10 min; (2) cell upper plate: the CTL action effect cell (1 × 10 of induction
5/ hole), the T2A2 cell of lotus peptide is as irritation cell (1 × 10
5/ hole) bed board.100 μ L/ holes.The distribution of cell in hole will try one's best evenly (after adding cell, do not shake again or bounce ELISPOT plate); ● positive control: cell concn is 1 × 10
5/ hole, add 10 μ L PHA, the secretion of this concentration energy effective stimulus IFN-γ; ● the negative contrast of background: RPMI 1640 substratum containing 5%FCS that add 100 μ L; (3) after adding all samples, cover plate lid, put into CO
2incubator, cultivates 18h for 37 ℃; (4) lysing cell: topple over cell and substratum in hole, 200 μ L/ holes add ice-cold deionized water, 4 ℃ of ice baths react 10 min (Low Osmotic Method lysing cell); (5) washing: topple over the liquid in hole, 1 × Washing Buffer, 200 μ L/ holes, washs 5 ~ 7 times, stop 30 ~ 60 s at every turn, last, buckle and do on thieving paper; (6) add detection antibody incubation: every hole adds the biotin labeling that 100 μ L have diluted to detect antibody, hatches 1h for 37 ℃; (7) washing: topple over the liquid in hole, 1 × Washing Buffer, 200 μ L/ holes, washs 5 times, each residence time is 30 ~ 60 s, last, buckles and do on thieving paper; (8) enzyme connection avidin is hatched: the enzyme connection avidin working fluid having diluted is joined to experimental port, and 100 μ L/ holes, hatch 1h for 37 ℃; (9) washing: topple over the liquid in hole, 1 × Washing Buffer, 200uL/ hole, wash 5 times, each residence time is 30 ~ 60 s, last, buckles and do on thieving paper; (10) colour developing: the AEC nitrite ion that thaws and prepared.Every hole adds the nitrite ion of 100 μ L, and room temperature leaves standstill 15-45min (at 20-25 ° of C, 25min is more suitable in colour developing), notes lucifuge; (11) color development stopping: topple over liquid in hole, open plate base, with deionized water wash 3 ~ 5 times, color development stopping process.Plate is tipped upside down on thieving paper, pat dry the tiny globule, take off afterwards protective layer, be placed on the place of ventilation, room temperature leaves standstill 10-30min, allows film naturally dry; Count the spot number of every hole formation in 96 orifice plates with ELISPOT Quantimet.
External ELISPOT experimental result shows: candidate's peptide P22, P57, P109, P129, P173 all can induce and obtain CTL in the peripheral blood of 6 health donors, and the CTL obtaining secretes the IFN-γ (as Fig. 7-11) of higher amount after stimulating.
The epitope peptide of the metastases related antigen that the present invention identifies provides theoretical basis for the tumor therapeutic polypeptide vaccine of development based on antigen MTA1, and lays a good foundation for the structure of the antigen peptide vaccine of follow-up multivalence, long peptide vaccine, polyepitope vaccines etc.
Application Project
-------------------
Antitumor CTL epitope peptide and the application thereof in <120> Title: MTA1 source
<130> AppFileReference :
<140> CurrentAppNumber : 201310112763.0
<141> CurrentFilingDate: on March 27th, 2013
Sequence
--------
<213> OrganismName: artificial sequence
<400> PreSequenceString :
tyrleuilea rgargilegl ugluleu 27
<212> Type : DNA/RNA
<211> Length : 9
SequenceName: aminoacid sequence P22
SequenceDescription : Tyr-Leu-Ile-Arg-Arg-Ile-Glu-Glu-Leu
Sequence
--------
<213> OrganismName: artificial sequence
<400> PreSequenceString :
alaleualaa splyshisal athrleu 27
<212> Type : DNA/RNA
<211> Length : 9
SequenceName: aminoacid sequence P57
SequenceDescription : Ala-Leu-Ala-Asp-Lys-His-Ala-Thr-Leu
Sequence
--------
<213> OrganismName: artificial sequence
<400> PreSequenceString :
pheleusera rgglnleugl userleu 27
<212> Type : DNA/RNA
<211> Length : 9
SequenceName: aminoacid sequence P109
SequenceDescription : Phe-Leu-Ser-Arg-Gln- Leu-Glu-Ser-Leu
Sequence
--------
<213> OrganismName: artificial sequence
<400> PreSequenceString :
thrleuleua sngluthrgl userleu 27
<212> Type : DNA/RNA
<211> Length : 9
SequenceName: aminoacid sequence P129
SequenceDescription : Thr- Leu-Leu-Asn-Glu-Thr-Glu-Ser-Leu
Sequence
--------
<213> OrganismName: artificial sequence
<400> PreSequenceString :
tyr-gln-ala-asp-ile-thr-asp-leu-leu
<212> Type : DNA/RNA
<211> Length : 9
SequenceName: aminoacid sequence P173
SequenceDescription : Tyr-Gln-Ala-Asp-Ile-Thr-Asp-Leu-Leu
Claims (2)
1. the antitumor CTL epitope peptide in MTA1 source, is made up of nine amino-acid residues, it is characterized in that: the aminoacid sequence of described epitope peptide is P22:Tyr-Leu-Ile-Arg-Arg-Ile-Glu-Glu-Leu.
2. the antitumor CTL epitope peptide in MTA1 claimed in claim 1 source is being prepared the application of tumor therapeutic polypeptide vaccine of this antigen of high expression level.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310112763.0A CN103214556B (en) | 2011-05-26 | 2011-05-26 | Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1) and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310112763.0A CN103214556B (en) | 2011-05-26 | 2011-05-26 | Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1) and application thereof |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110138635 Division CN102212112B (en) | 2011-05-26 | 2011-05-26 | MTA1 (metastasis-associated gene 1)-derived anti-tumor CTL (cytotoxic T lymphocyte) epitope peptide and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103214556A CN103214556A (en) | 2013-07-24 |
| CN103214556B true CN103214556B (en) | 2014-06-04 |
Family
ID=48812761
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310112763.0A Active CN103214556B (en) | 2011-05-26 | 2011-05-26 | Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1) and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103214556B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110577580B (en) * | 2019-09-25 | 2023-03-24 | 成都奥达生物科技有限公司 | Multiple myeloma treatment vaccine |
-
2011
- 2011-05-26 CN CN201310112763.0A patent/CN103214556B/en active Active
Non-Patent Citations (6)
| Title |
|---|
| Geng Li et al..Identification of metastasis associated antigen 1(MTA1) by serological screening of prostate cancer cDAN libraries.《The Open Biochemistry Journal》.2008, |
| Identification of metastasis associated antigen 1(MTA1) by serological screening of prostate cancer cDAN libraries;Geng Li et al.;《The Open Biochemistry Journal》;20081231;全文 * |
| MTA1基因与肿瘤转移相关性的研究进展;高玉军;《山东医药》;20101231;全文 * |
| NP_001068982;Smith,T.P.et al.;《GenBank》;20110413 * |
| Smith,T.P.et al..NP_001068982.《GenBank》.2011, |
| 高玉军.MTA1基因与肿瘤转移相关性的研究进展.《山东医药》.2010, |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103214556A (en) | 2013-07-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105777911B (en) | Anti- BCMA Chimeric antigen receptor, encoding gene, recombinant expression carrier and its construction method and application | |
| CN105296431B (en) | The α β T cells and its suppression cancer purposes of tumor combination specificity gamma delta T CR genetic modifications | |
| CN105820255B (en) | Anti-CD 33 Chimeric antigen receptor, encoding gene, recombinant expression carrier and its construction method and application | |
| CN105949318A (en) | Anti-HER2 chimeric antigen receptor, encoding gene, recombinant expression vector, construction method of recombinant expression vector, and application | |
| CN105949317A (en) | Anti-CD20 chimeric antigen receptor, encoding gene, recombinant expression vector, construction method of recombinant expression vector, and application | |
| CN106589133A (en) | Preparation and applications of liver-cancer-specific CTL cells induced by new enhanced antigen combined polypeptide | |
| CN105950663A (en) | CD 30-targeting replication-defective recombinant lentivirus CAR-T transgenic vector as well as construction method and applications thereof | |
| CN105950662B (en) | A kind of replication defective recombinant slow virus CAR-T transgene carrier targeting CD22 and its construction method and application | |
| CN103214556B (en) | Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MAT1 (Metastasis Associated Gene1) and application thereof | |
| CN103204908B (en) | MTA1-derived anti-tumour CTL (cytotoxiclymphocyte) epitope peptide and application thereof | |
| CN103145801B (en) | Anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide from MTA1 (Tumor Metastasis Associated Antigen 1) and application thereof | |
| CN102212112B (en) | MTA1 (metastasis-associated gene 1)-derived anti-tumor CTL (cytotoxic T lymphocyte) epitope peptide and application thereof | |
| CN106589106A (en) | Specific CTL epitope peptide of HLA-A24 restricted ECM1 and application thereof | |
| CN102516363B (en) | CTL (Cytotoxic T Lymphocytes) epitope peptide from PL2L60 source and application thereof | |
| Qin et al. | Melanoma B16-F1 cells coated with fusion protein of mouse calreticulin and virus G-protein coupled receptor induced the antitumor immune response in Balb/C mice | |
| CN109371005A (en) | HLA-0201 restrictive PADI4 epitope polypeptide and application thereof | |
| CN106117339A (en) | The CTL of a kind of specific tumor antigen NSE identifies epitope peptide and application thereof | |
| CN102153624A (en) | Antitumor cytotoxic T lymphocyte (CTL) epitope peptide analog derived from COX-2 | |
| CN101870724B (en) | PLAC1 (Placenta-specific 1) anti-tumor CTL (Cytotoxic T Lymphocyte) epitope peptide and application thereof | |
| CN106117334B (en) | CTL recognition epitope peptide of tumor antigen MAGE3 and application thereof | |
| CN106008692A (en) | CTL identification epitope peptide of tumor antigen GPC3 and application of epitope peptide | |
| CN104374918B (en) | Heat shock protein 47 glioma associated tumor antigen and application | |
| CN106117337B (en) | CTL recognition epitope peptide of specific tumor antigen SF and application thereof | |
| CN101948508B (en) | Tumor-resistance CTL (Cytotoxic T Lymphocyte) epitope peptide derived from PIWIL2 and application thereof | |
| CN104761636B (en) | A kind of restricted eEF2 epitope polypeptides of HLA-A33 and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230803 Address after: 450000 building 11, Zhengzhou airport biomedical park, northeast corner of the intersection of Huanghai Road and biotechnology Second Street, Hangkong port area, Zhengzhou City, Henan Province Patentee after: Zhengzhou Yuanchuang Gene Technology Co.,Ltd. Address before: 450001 No. 100 science Avenue, hi tech Zone, Henan, Zhengzhou Patentee before: Zhengzhou University |
|
| TR01 | Transfer of patent right |