CN101717449A - Recombinant TRAIL-Fc fusion protein as well as preparation and application thereof - Google Patents
Recombinant TRAIL-Fc fusion protein as well as preparation and application thereof Download PDFInfo
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Abstract
The invention relates to a design technology for fusion protein and a protein expression and purification technology, and relates the application of fusion protein as a tumor therapeutic medicament. The invention is characterized by fusing the 114th-281th amino acid of tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) with human IgG1Fc. The fused protein consists of 400 amino acids, the isoelectric point pI is 9.34, and theoretical molecular weight is 45535.30. The protein exists by a dimer or a trimer, can remarkably induce apoptosis of a plurality of kinds of tumor cells and killsor/and inhibit tumor cells in an animal body. Compared with TRAIL, the TRAIL-Fc fusion protein has large molecular weight, high activity and long vivo half-life, is expressed by mammalian cells, and can be used as a tumor therapeutic medicament.
Description
Technical field
The invention belongs to biological technical field, particularly TRAIL-Fc fusion rotein designing technique and protein expression and purification technology relate to fusion rotein as the application in the anti-tumor medicine.
Background technology
TRAIL (TNF-Related Apoptosis Inducing Ligand), promptly tumor necrosis factor relative cell death inducing ligand is a member in the TNF superfamily.People TRAIL is an II type transmembrane protein, and promptly its C end is positioned at outside the film, and the N end is positioned at born of the same parents.TRAIL is made up of 281 amino acid, and molecular weight is 32KD, and its extracellular region is made up of 243 amino acid.The physiological function of TRAIL is a cell death inducing, though TRAIL all has expression at many normal cells such as myocardial cell, lymphocyte, splenocyte, prostatic cell, gonad cell and small intestine cells etc., but it is inducing tumor cell and viral cell transformed apoptosis only, comprises intestinal cancer, mammary cancer, melanoma, lung cancer, kidney, central nerve neuroma, leukemia, skin carcinoma and multiple myeloma cells etc.Discover that the TRAIL extracellular region can be exercised its physiological function.By further investigation to its extracellular region, find many brachymemmas extracellular region also have the cell such as the TRAIL of cell death inducing
95-281, TRAL
112-281, TRAIL
114-281Deng, present TRAIL
114-281(AMG655) carry out clinical trial by U.S. Amgen company.
But because the transformation period of TRAIL in blood plasma had only about 30 minutes, the cancer patients needs administration every day and dosage big (15mg/kg/d), so be necessary to prolong its transformation period in blood plasma.Reorganization TRAIL all adopts the coli expression system preparation at present, and expression product is the polymer form, needs just to have short apoptosis of tumor cells effect after the renaturation.
Human IgG1 Fc is hinge region (Hinge) and complement land (CH2, the CH3) of heavy chain of antibody.In the exploitation of recombinant products, usually itself and target protein are merged to increase the transformation period of target protein.It promptly is a kind of soluble fusion protein by people's cytotoxic T cell antigen 4 (CTLA4) extracellular domain and human normal immunoglobulin IgG1 (IgG1) Fc section two portions be combined into that the first selectivity of getting permission to go on the market stimulates conditioning agent class biotechnological formulation Ah crust former times general (Abatacept) (100 o'clock Shi Guibao companies) altogether.After having incorporated IgG1Fc, the transformation period reached for two weeks.
Proved that TRAIL plays a role with its homology two, trimerical form.Because have three halfcystines at the IgG1Fc hinge region, IgG1Fc very easily forms dimer, after merging with it, target protein forms dimer or polymer easily.TRAIL target protein and IgG1Fc merge the artifact activity and can be improved, and the transformation period can significant prolongation.
Summary of the invention
The invention reside in provides a kind of dimerization or tripolymer people TRAIL-Fc fusion rotein with big, the active height of molecular weight and long half time.
Another object of the present invention is to provide a kind of method of producing this fusion rotein, described method is a Mammals CHO-k1 cell expressing.The TRAIL-Fc fusion rotein that this method produces is dimerization or tripolymer, and its purification process is simple, the purity height, and biological activity is strong.
Another object of the present invention is to provide this fusion rotein as the application in the anti-tumor medicine.
Fusion rotein provided by the invention is one and contains 400 amino acid whose albumen, this albumen iso-electric point pI=8.34, and theoretical molecular is 45535.30, its aminoacid sequence is Seq1.Being TRAIL114-281 from the 1-168 position in the TRAIL-Fc sequence, is transformed IgG1Fc hinge region from the 169-183 position, is the CH2 district of IgG1Fc from the 184-293 position, is the CH3 district of IgG1Fc from the 294-400 position.
Technical scheme:
1, makes up mammalian cell expression recombinant plasmid pSecTag2A-TRAIL-Fc.
2, recombinant plasmid transformed intestinal bacteria TOP10, screening positive clone, the plasmid that extracts is identified, is checked order and identify with HindIII and EcoRI double digestion.The result proves: resulting positive colony and design in full accord.
3, it is standby that middle amount is extracted recombinant plasmid pSecTag2A-TRAIL-Fc.
4, recombinant plasmid is used liposome method transient transfection HEK293T cell, 37 ℃, 5%CO
2Cultivate and collect culture supernatant after three days.With DotBlot and the proteic expression of Western Blot testing goal.
5, recombinant plasmid is used calcium phosphate method transfection CHO-k1 cell, with the Zeocine of the 250 μ m/mL screening of pressurizeing, use the DotBlot screening positive clone, positive colony is carried out enlarged culturing, after Western Blot identifies, culture supernatant is carried out nickel post affinitive layer purification, and the enteropeptidase enzyme is cut, Protein A post affinitive layer purification.
6, the gained sample carries out the inducing apoptosis of tumour cell experiment.
7, the gained sample carries out the experiment of tumor-bearing mice tumor suppression.
Description of drawings
The structure of accompanying drawing 1 recombinant expression plasmid, goal gene are that carrier is all behind the HindIII+EcoRI double digestion, with the connection of T4 dna ligase.
The enzyme of accompanying drawing 2 recombinant expression plasmids is cut evaluation.1:pSectag2A, 2,3,4: cloned plasmids to be measured, M:DNA Marker.Recombinant plasmid is behind the HindIII+EcoRI double digestion, and 3, No. 4 a 1.2kb band and a 5.2kb band appear in cloned plasmids, and the 1.2kb band is a goal gene, and the 5.2kb band is a carrier segments, and the result shows that construction of recombinant plasmid is correct.
Accompanying drawing 3DotBlot experimental result will be checked on the transient expression in pvdf membrane, does one with mouse anti His-Tag and resists, and goat anti-mouse IgG-HRP does two and resists chemical luminous substrate development Kodak egative film." E " is the transient expression result, "+" positive contrast, "-" negative contrast.The result shows has target protein to express.
Accompanying drawing 4Western Blot experiment will be checked on the transient expression in pvdf membrane, does one with mouse anti His-Tag and resists, and goat anti-mouse IgG-HRP does two and resists chemical luminous substrate development Kodak egative film.1: sample is by non-reduced processing; 2: sample is handled by reduction; 3: protein molecular weight standard.The result shows under the target protein nature situation and exists with the polymer form that exist with monomeric form, molecular weight and theoretical molecular coincide under the reduction situation.
Accompanying drawing 5PBS, TRAIL
114-281Compare with the tumour size after the TRAIL-Fc processing of the present invention, TRAIL-Fc of the present invention injects the growth that once can significantly suppress the mouse interior tumor cell, P＜0.01 weekly.
Embodiment
For the ease of purifying, add label H is * 6-GGGS-DDDDK in the front of TRAIL-Fc, GGGS is a connection peptides, DDDDK is that enteropeptidase is cut the site, thereby forms His * 6-GGGS-DDDDK-TRAIL
114-281-Fc sequence.The front and back restriction enzyme site of cDNA sequence is respectively HindIII and EcoRI.This sequence adopts artificial complete synthesis mode synthetic, and its sequence is seen Seq4.Seq4 and pSecTag2A behind HindIII and EcoRI double digestion, are connected and composed pSecTag2A-TRAIL-Fc, see accompanying drawing 1.Recombinant plasmid transformed intestinal bacteria TOP10, the picking positive colony is cultivated in containing the LB substratum of penbritin, extracts plasmid and does the evaluation of HindIII+EcoRI double digestion, sees accompanying drawing 2.Further carrying out dna sequencing identifies.
1.TRAIL-Fc transient expression.With recombinant plasmid liposome method transient transfection HEK293T cell, 37 ℃, 5%CO
2Cultivate and collect culture supernatant after three days.Make one anti-ly of mouse anti His-Tag antibody, goat anti-mouse IgG-HRP does two and anti-ly carries out DotBlot and Western Blot detects, and to judge the expression of target protein, the results are shown in accompanying drawing 3.
2.TRAIL-Fc expression at the CHO-k cell.Determine after the Zeocine tolerance concentration of CHO-k cell with calcium phosphate method transfection recombinant plasmid to the CHO-k cell, add the Zeocine screening, the substratum of drawing in the corresponding cell clone is done DotBlot and Western Blot detection, get positive colony and be amplification culture to 10 layer CellFactory step by step, cultivate with the DMEM that does not contain glutamine and to use serum free medium after a week instead and continue to cultivate for 4 weeks, collect supernatant.
Culture supernatant is concentrated, replaces damping fluid to 10mM Tris with ultrafiltration process, 100mM NaCl (pH 7.2); Ni-Chelating Sepharose FF chromatography column is gone up sample through 10mM Tris after 100mM NaCl (pH 7.2) balance, reequilibrate is to baseline; With containing the 100mM imidazoles, 10mM Tris, 100mM NaCl (pH7.2) buffer solution elution target protein.The albumen elutriant adds enteropeptidase (1: 500 mass ratio) digestion, enzyme is cut the back sample through Protein A-Agarose chromatography column (10mM PB buffering, pH 7.4) absorption after, with 0.1M Gly (pH 3.0) buffer solution elution purpose, add 2M Tris (pH 7.2) (about 1/10 elution volume of total amount) while receiving when collecting the purpose peak and neutralize.Dialysis target protein damping fluid is to PBS.
Embodiment 4, and TRAIL-Fc kills and wounds the tumor cell in vitro experiment
With the TRAIL-Fc of purifying by 100,25,6.25,1.56,0.39 and the concentration of 0.09ng/mL handle various tumor cell line, as Hela clone (human cervical carcinoma cell), KU-8 clone (people's penile cancer cell), U937 clone (human medullary leukemia cell), IM-9 clone (human myeloma cell), A549 clone (human lung carcinoma cell), MCF-7 clone (human breast cancer cell) and Jurkat clone (human T lymphocyte leukemia cell), act on after 48 hours and dyeing, measure the 570nm absorption value with mtt assay.The result proves that TRAIL-Fc has the effect of killing tumor cell widely, and wherein, Jurkat clone and MCF-7 clone are the highest to the susceptibility of TRAIL-Fc.The TRAIL-Fc protein concentration of inducing Jurkat cell 50% apoptosis rate is about 2ng/mL.
Table 1TRAIL-Fc is to the maximal percentage inhibition of various tumour cells
Embodiment 5, and TRAIL-Fc kills and wounds the mouse interior tumor cell experiment
Animal for research: the Balb/C mouse, totally 18, be divided into three groups, 6 every group, one group is the PBS negative control, one group is TRAIL
114-281Treatment group, another group TRAIL-Fc treatment group.Mouse S180 oncocyte (4 * 10 with vitro culture
6/ only) the back subcutaneous injection, simultaneously, subcutaneous injection TRAIL
114-281(100 μ g/ only), TRAIL-Fc (234 μ g/, mol ratio equates) or PBS.TRAIL-Fc injects weekly once, TRAIL
114-281Be administered once every day.4 weeks back execution mouse is measured the size and the weight that form tumour, and the result shows that all there is tumor growth at the back of PBS group, and is evenly big or small; TRAIL
114-281The group mouse has not bearing tumor of three backs, and two tumours are less, and a tumour is bigger; TRAIL-Fc group mouse has not bearing tumor of three backs, and three less.Discovery TRAIL-Fc is administered once weekly and can significantly suppresses the growth of mouse interior tumor cell, P＜0.01 after the statistical procedures.Prove that TRAIL-Fc of the present invention has long-acting antitumor action in the body.
Sequence table
＜110〉Chongqing Fujin Biological Medicine Co. Ltd
＜120〉reorganization TRAIL-Fc fusion rotein and preparation and application
<130>a
<160>3
<170>PatentIn?version?3.3
<210>1
<211>400
<212>PRT
＜213〉people
<400>1
Val?Arg?Glu?Arg?Gly?Pro?Gln?Arg?Val?Ala?Ala?His?Ile?Thr?Gly?Thr
1 5 10 15
Arg?Gly?Arg?Ser?Asn?Thr?Leu?Ser?Ser?Pro?Asn?Ser?Lys?Asn?Glu?Lys
20 25 30
Ala?Leu?Gly?Arg?Lys?Ile?Asn?Ser?Trp?Glu?Ser?Ser?Arg?Ser?Gly?His
35 40 45
Ser?Phe?Leu?Ser?Asn?Leu?His?Leu?Arg?Asn?Gly?Glu?Leu?Val?Ile?His
50 55 60
Glu?Lys?Gly?Phe?Tyr?Tyr?Ile?Tyr?Ser?Gln?Thr?Tyr?Phe?Arg?Phe?Gln
65 70 75 80
Glu?Glu?Ile?Lys?Glu?Asn?Thr?Lys?Asn?Asp?Lys?Gln?Met?Val?Gln?Tyr
85 90 95
Ile?Tyr?Lys?Tyr?Thr?Ser?Tyr?Pro?Asp?Pro?Ile?Leu?Leu?Met?Lys?Ser
100 105 110
Ala?Arg?Asn?Ser?Cys?Trp?Ser?Lys?Asp?Ala?Glu?Tyr?Gly?Leu?Tyr?Ser
115 120 125
Ile?Tyr?Gln?Gly?Gly?Ile?Phe?Glu?Leu?Lys?Glu?Asn?Asp?Arg?Ile?Phe
130 135 140
Val?Ser?Val?Thr?Asn?Glu?His?Leu?Ile?Asp?Met?Asp?His?Glu?Ala?Ser
145 150 155 160
Phe?Phe?Gly?Ala?Phe?Leu?Val?Gly?Glu?Pro?Lys?Ser?Cys?Asp?Lys?Thr
165 170 175
His?Thr?Ser?Pro?Pro?Ser?Pro?Ala?Pro?Glu?Leu?Leu?Gly?Gly?Pro?Ser
180 185 190
Val?Phe?Leu?Phe?Pro?Pro?Lys?Pro?Lys?Asp?Thr?Leu?Met?Ile?Ser?Arg
195 200 205
Thr?Pro?Glu?Val?Thr?Cys?Val?Val?Val?Asp?Val?Ser?His?Glu?Asp?Pro
210 215 220
Glu?Val?Lys?Phe?Asn?Trp?Tyr?Val?Asp?Gly?Val?Glu?Val?His?Asn?Ala
225 230 235 240
Lys?Thr?Lys?Pro?Arg?Glu?Glu?Gln?Tyr?Asn?Ser?Thr?Tyr?Arg?Val?Val
245 250 255
Ser?Val?Leu?Thr?Val?Leu?His?Gln?Asp?Trp?Leu?Asn?Gly?Lys?Glu?Tyr
260 265 270
Lys?Cys?Lys?Val?Ser?Asn?Lys?Ala?Leu?Pro?Ala?Pro?Ile?Glu?Lys?Thr
275 280 285
Ile?Ser?Lys?Ala?Lys?Gly?Gln?Pro?Arg?Glu?Pro?Gln?Val?Tyr?Thr?Leu
290 295 300
Pro?Pro?Ser?Arg?Asp?Glu?Leu?Thr?Lys?Asn?Gln?Val?Ser?Leu?Thr?Cys
305 310 315 320
Leu?Val?Lys?Gly?Phe?Tyr?Pro?Ser?Asp?Ile?Ala?Val?Glu?Trp?Glu?Ser
325 330 335
Asn?Gly?Gln?Pro?Glu?Asn?Asn?Tyr?Lys?Thr?Thr?Pro?Pro?Val?Leu?Asp
340 345 350
Ser?Asp?Gly?Ser?Phe?Phe?Leu?Tyr?Ser?Lys?Leu?Thr?Val?Asp?Lys?Ser
355 360 365
Arg?Trp?Gln?Gln?Gly?Asn?Val?Phe?Ser?Cys?Ser?Val?Met?His?Glu?Ala
370 375 380
Leu?His?Asn?His?Tyr?Thr?Gln?Lys?Ser?Leu?Ser?Leu?Ser?Pro?Gly?Lys
385 390 395 400
<210>2
<211>15
<212>PRT
＜213〉synthetic
<400>2
Glu?Pro?Lys?Ser?Cys?Asp?Lys?Thr?His?Thr?Ser?Pro?Pro?Ser?Pro
1 5 10 15
<210>3
<211>15
<212>PRT
＜213〉synthetic
<400>3
His?His?His?His?His?His?Gly?Gly?Gly?Ser?Asp?Asp?Asp?Asp?Lys
1 5 10 15
Nucleic acid and aminoacid sequence Chinese edition .ST25 in the reorganization TRAILFc patent
His?His?His?His?His?His?Gly?Gly?Gly?Ser?Asp?Asp?Asp?Asp?Lys
1 5 10 15
Claims (9)
1. a reorganization TRAIL-Fc albumen is characterized in that this albumen is a fusion rotein, by TRAIL
114-281The CH2 of aminoacid sequence and human IgG1 Fc and CH3 fragment merge.
2. the described fusion rotein of claim 1 is characterized in that, this aminoacid sequence is Seq1:
VAL?ARG?GLU?ARG?GLY?PRO?GLN?ARG?VAL?ALA?ALA?HIS?ILE?THR?GLY?THR
ARG?GLY?ARG?SER?ASN?THR?LEU?SER?SER?PRO?ASN?SER?LYS?ASN?GLU?LYS
ALA?LEU?GLY?ARG?LYS?ILE?ASN?SER?TRP?GLU?SER?SER?ARG?SER?GLY?HIS
SER?PHE?LEU?SER?ASN?LEU?HIS?LEU?ARG?ASN?GLY?GLU?LEU?VAL?ILE?HIS
GLU?LYS?GLY?PHE?TYR?TYR?ILE?TYR?SER?GLN?THR?TYR?PHE?ARG?PHE?GLN
GLU?GLU?ILE?LYS?GLU?ASN?THR?LYS?ASN?ASP?LYS?GLN?MET?VAL?GLN?TYR
ILE?TYR?LYS?TYR?THR?SER?TYR?PRO?ASP?PRO?ILE?LEU?LEU?MET?LYS?SER
ALA?ARG?ASN?SER?CYS?TRP?SER?LYS?ASP?ALA?GLU?TYR?GLY?LEU?TYR?SER
ILE?TYR?GLN?GLY?GLY?ILE?PHE?GLU?LEU?LYS?GLU?ASN?ASP?ARG?ILE?PHE
VAL?SER?VAL?THR?ASN?GLU?HIS?LEU?ILE?ASP?MET?ASP?HIS?GLU?ALA?SER
PHE?PHE?GLY?ALA?PHE?LEU?VAL?GLY?GLU?PRO?LYS?SER?CYS?ASP?LYS?THR
HIS?THR?SER?PRO?PRO?SER?PRO?ALA?PRO?GLU?LEU?LEU?GLY?GLY?PRO?SER
VAL?PHE?LEU?PHE?PRO?PRO?LYS?PRO?LYS?ASP?THR?LEU?MET?ILE?SER?ARG
THR?PRO?GLU?VAL?THR?CYS?VAL?VAL?VAL?ASP?VAL?SER?HIS?GLU?ASP?PRO
GLU?VAL?LYS?PHE?ASN?TRP?TYR?VAL?ASP?GLY?VAL?GLU?VAL?HIS?ASN?ALA
LYS?THR?LYS?PRO?ARG?GLU?GLU?GLN?TYR?ASN?SER?THR?TYR?ARG?VAL?VAL
SER?VAL?LEU?THR?VAL?LEU?HIS?GLN?ASP?TRP?LEU?ASN?GLY?LYS?GLU?TYR
LYS?CYS?LYS?VAL?SER?ASN?LYS?ALA?LEU?PRO?ALA?PRO?ILE?GLU?LYS?THR
ILE?SER?LYS?ALA?LYS?GLY?GLN?PRO?ARG?GLU?PRO?GLN?VAL?TYR?THR?LEU
PRO?PRO?SER?ARG?ASP?GLU?LEU?THR?LYS?ASN?GLN?VAL?SER?LEU?THR?CYS
LEU?VAL?LYS?GLY?PHE?TYR?PRO?SER?ASP?ILE?ALA?VAL?GLU?TRP?GLU?SER
ASN?GLY?GLN?PRO?GLU?ASN?ASN?TYR?LYS?THR?THR?PRO?PRO?VAL?LEU?ASP
SER?ASP?GLY?SER?PHE?PHE?LEU?TYR?SER?LYS?LEU?THR?VAL?ASP?LYS?SER
ARG?TRP?GLN?GLN?GLY?ASN?VAL?PHE?SER?CYS?SER?VAL?MET?HIS?GLU?ALA
LEU?HIS?ASN?HIS?TYR?THR?GLN?LYS?SER?LEU?SER?LEU?SER?PRO?GLY?LYS
3. the described fusion rotein of claim 1, it is characterized in that first amino acid of its hinge region of human IgG1 Fc sports L-glutamic acid Glu by Xie Ansuan Val, only keep a halfcystine Cys in whole hinge area, its latter two Cys substitutes with Ser respectively, and its hinge region sequence is Seq2:
GLU?PRO?LYS?SER?CYS?ASP?LYS?THR?HIS?THR?SER?PRO?PRO?SER?PRO
4. the described fusion rotein of claim 1, in expressing design, purification tag His * 6, connection peptides GGGS and protease cutting site DDDDK are introduced in the maturation protein front, and its sequence is Seq3:
HIS?HIS?HIS?HIS?HIS?HIS?GLY?GLY?GLY?SER?ASP?ASP?ASP?ASP?LYS
5. the pairing nucleotide sequence of the described fusion rotein of claim 1.
6. the described fusion rotein of claim 1 is applicable in intestinal bacteria, yeast and mammalian cell and expresses.
7. the expression of the described fusion rotein of claim 1 in the mammalian cell Chinese hamster ovary celI.
8. the expression supernatant in the claim 7 is cut purifying process with Protein A affinity chromatography through ultrafiltration, nickel affinity chromatography, enteropeptidase enzyme.
9. the described application of fusion rotein in the preparation anti-tumor medicine of claim 1.
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| CN102775497A (en) * | 2012-07-13 | 2012-11-14 | 浙江大学 | TNF (Tumor Necrosis Factor)-related apoptosis-inducing ligand fusion protein and preparation method thereof |
| WO2013037090A1 (en) * | 2011-09-16 | 2013-03-21 | 北京沙东生物技术有限公司 | Fusion protein comprising circularly permuted form of trail/apo2l, coding gene and use thereof |
| WO2016127346A1 (en) * | 2015-02-11 | 2016-08-18 | 四川大学华西医院 | Tumour necrosis factor-related apoptosis-inducing ligand variant and preparation method and use thereof |
| CN106046170A (en) * | 2015-04-10 | 2016-10-26 | 中国医学科学院药物研究所 | Novel TRAIL fusion protein |
| CN106978429A (en) * | 2017-03-16 | 2017-07-25 | 中国人民解放军第四军医大学 | A kind of bovine enterokinase light chain load magnetic bead and its preparation method and application |
| CN108642070A (en) * | 2018-04-11 | 2018-10-12 | 沈阳金石生物制药有限公司 | Recombined human Fc antibody of specific inducing apoptosis of tumour cell and preparation method thereof, purposes |
| CN110386985A (en) * | 2018-04-19 | 2019-10-29 | 四川大学华西医院 | A kind of tumor-targeting promotees apoptotic fusion proteins and application thereof |
| CN113717289A (en) * | 2021-07-23 | 2021-11-30 | 华东理工大学 | SAC-TRAIL fusion protein and preparation method and application thereof |
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| CN103534273A (en) * | 2011-09-16 | 2014-01-22 | 北京沙东生物技术有限公司 | Fusion protein comprising circularly permuted form of TRAIL/APO2L, coding gene and use thereof |
| WO2013037090A1 (en) * | 2011-09-16 | 2013-03-21 | 北京沙东生物技术有限公司 | Fusion protein comprising circularly permuted form of trail/apo2l, coding gene and use thereof |
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| CN106046170A (en) * | 2015-04-10 | 2016-10-26 | 中国医学科学院药物研究所 | Novel TRAIL fusion protein |
| CN106046170B (en) * | 2015-04-10 | 2020-07-10 | 中国医学科学院药物研究所 | Novel TRAI L fusion protein |
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| CN108642070A (en) * | 2018-04-11 | 2018-10-12 | 沈阳金石生物制药有限公司 | Recombined human Fc antibody of specific inducing apoptosis of tumour cell and preparation method thereof, purposes |
| CN108642070B (en) * | 2018-04-11 | 2022-03-15 | 沈阳金石生物制药有限公司 | Recombinant human Fc antibody for specifically inducing tumor cell apoptosis and preparation method and application thereof |
| CN110386985A (en) * | 2018-04-19 | 2019-10-29 | 四川大学华西医院 | A kind of tumor-targeting promotees apoptotic fusion proteins and application thereof |
| CN113717289A (en) * | 2021-07-23 | 2021-11-30 | 华东理工大学 | SAC-TRAIL fusion protein and preparation method and application thereof |
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