CN110684113A - Bispecific antibody for resisting PD-1 antigen and c-Met antigen and construction method thereof - Google Patents
Bispecific antibody for resisting PD-1 antigen and c-Met antigen and construction method thereof Download PDFInfo
- Publication number
- CN110684113A CN110684113A CN201810748597.6A CN201810748597A CN110684113A CN 110684113 A CN110684113 A CN 110684113A CN 201810748597 A CN201810748597 A CN 201810748597A CN 110684113 A CN110684113 A CN 110684113A
- Authority
- CN
- China
- Prior art keywords
- met
- antigen
- bispecific antibody
- antibody
- heavy chain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
- C07K16/2803—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
- C07K16/2818—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 CD28 or CD152
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of biology, and particularly relates to a recombinant humanized PD-1 and c-Met bispecific antibody and a construction method thereof. On the basis of computer molecular structure simulation, the invention utilizes molecular cloning technology to take two screened monoclonal antibodies as templates to construct expression plasmids of the bispecific antibody, the expression plasmids are tetravalent bispecific antibodies, can recognize c-Met on the surface of tumor cells and block the combination of negative co-stimulation molecules PD-1 on the surface of T cells and PD-L1 on the surface of the tumor cells, so that the T cells generate continuous killing effect on the tumor cells and provide a foundation for immune intervention of targeting c-Met positive tumor cells.
Description
Technical Field
The invention belongs to the technical field of biology, relates to a bispecific antibody and a construction method thereof, and particularly relates to a recombinant humanized PD-1& c-Met bispecific antibody and a construction method thereof.
Background
The prior art discloses that T cells are the core of the anti-tumor immune response of the immune system of the body, and the complete activation of the T cells depends on the combined action of double signals and cytokines. The programmed cell death receptor (PD-1) belongs to negative co-stimulation molecules in a second signal required by T cell activation, and can down-regulate the activity of CD3(CD28) induced phosphatidylinositol 3 kinase and protein kinase B after being combined with ligands PD-L1 and PD-L2 thereof, inhibit the transcription of T cell proliferation related genes and promote the apoptosis of the T cell proliferation related genes; meanwhile, Protein Tyrosine Phosphatase 1 (PTP-1) is recruited, so that the combined molecules are dephosphorylated, and the proliferation and activation of T cells are inhibited, thereby reducing the immune response of the T cells and being beneficial to maintaining the autoimmune tolerance; studies have shown that tumor cells also take advantage of this point by expressing PD-L1 on their surface to silence T cell function and thereby escape immune surveillance.
The targeted therapy of tumor is to design corresponding antitumor drug aiming at the difference of the surface molecular expression of tumor and normal tissue on the cellular molecular level, the drug enters into the body to kill tumor cells specifically without affecting the surrounding normal tissue, and the toxic and side effects generated by the traditional radiotherapy, chemotherapy and other therapeutic measures can be reduced to the maximum extent.
The Hepatocyte Growth Factor Receptor (HGFR), also known as c-Met, is encoded by proto-oncogene c-Met, has tyrosine kinase activity, can promote proliferation and differentiation of cells after being combined with ligand HGF thereof, and researches show that the c-Met is closely related to the occurrence and metastasis of various tumors, and the overactivation of the c-Met is found in a plurality of tumor patients, but has almost no activity in normal tissues.
The traditional monoclonal antibody generally causes apoptosis of cells by combining antigens on the surface of tumor cells, which are different from normal cells, and can also activate a complement system to achieve the purpose of killing the tumor cells, but the application of the traditional monoclonal antibody is limited due to the defect that T cells cannot be recruited to resist tumors.
Bispecific antibodies (BsAb), which can specifically bind to two different antigens simultaneously, were developed as early as 30 years ago and were not studied because of limited production technology and failed clinical trials; the development of genetic engineering technology in recent years provides a new opportunity for the development of humanized bispecific antibodies with lower immunogenicity, and the bispecific antibodies are mainly characterized by simultaneously combining tumor cell surface antigens and immune effector cell surface antigens, blocking the signal transmission of double targets and directly triggering the specific killing effect of immune effector cells on tumor cells.
Most bispecific antibodies currently under investigation select CD3 on the surface of T cells as a target on effector cells, and several studies show that after treatment with CD3 double antibody, the expression level of PD-1 on the surface of T cells and PD-L1 on the surface of tumor cells is increased, and the continuous activation of PD-1/PD-L1 pathway inhibits the function of T cells, thereby weakening the effect of killing tumor cells; at present, the commonly used bispecific antibody with asymmetric structure has the following defects, for example, the light and heavy chain mismatching phenomenon often occurs in self-assembly, while the small bispecific antibody has unstable structure and lacks Fc region, which results in short half-life period, easy elimination in vivo, and difficulty in achieving the purpose of effectively treating tumor, etc.
Based on the current situation of the prior art, the inventors of the present application intend to provide a bispecific antibody against PD-1 antigen and c-Met antigen, which can overcome the problems of insufficient therapeutic effect in tumor treatment caused by immunosuppression due to single recognition or after recognition, and the problems of unstable antibody structure or low assembly efficiency in the prior art.
Disclosure of Invention
The invention aims to provide a bispecific antibody of an anti-PD-1 antigen and an anti-c-Met antigen aiming at the problems in the prior art, in particular to a tetravalent bispecific antibody which can simultaneously combine a tumor-related antigen c-Met and an immune checkpoint molecule PD-1 and is combined with an anti-c-Met antibody to give a PD-1 inhibitor the capability of targeting c-Met positive tumor cells, thereby reducing the off-target probability and correspondingly reducing the side effect on normal tissues, and the bispecific antibody can block a PD-1/PD-L1 inhibitory pathway at any time based on the problems in the prior art that the curative effect is insufficient in tumor treatment and the antibody structure is unstable or the assembly efficiency is low, the sustained effect of killing tumor cells is generated.
In particular, the method comprises the following steps of,
the present invention provides a bispecific antibody against a PD-1 antigen and an anti c-Met antigen, comprising: a first antibody segment specifically recognizing the PD-1 antigen of T lymphocytes and a second antibody segment specifically recognizing the c-Met antigen of tumor cells.
In the present invention, the light chain unit of the bispecific antibody is: the variable region and the constant region of the light chain of the anti-PD-1 antibody, and the heavy chain unit of the bispecific antibody is: a C-Met scFv region is formed by a light chain variable region and a heavy chain variable region of the anti-C-Met antibody through a Gly4Ser connecting peptide, and the scFv region is connected to the C terminal of the heavy chain unit of the anti-PD-1 antibody through a Gly4Ser connecting peptide sequence to form the heavy chain unit of the bispecific antibody;
in the present invention, the nucleotide sequence of the light chain unit of the anti-PD-1 antibody is as shown in SEQ ID NO: 1, and the nucleotide sequence of the heavy chain unit of the anti-PD-1 antibody is shown as SEQ ID NO: 2, the nucleotide sequence of the light chain variable region of the anti-c-Met antibody is shown as SEQ ID NO: 3, the nucleotide sequence of the heavy chain variable region of the anti-c-Met antibody is shown as SEQ ID NO: 4, the nucleotide sequence of the Gly4Ser connecting peptide is shown as SEQ ID NO: 5, respectively.
The bispecific antibody against the PD-1 antigen and the c-Met antigen of the present invention is prepared by the following method:
(1) constructing the heavy chain and the light chain into the same expression vector respectively;
(2) co-transfecting the two expression vectors constructed in (1) together into a host cell, culturing and separating a supernatant;
(3) and purifying the separated supernatant to obtain the PD-1& c-Met bispecific antibody IgG-scFv.
In the invention, the expression vector is pCEP 4.
In the invention, the host cell is human embryonic kidney 293 cell HEK293E modified by EBNA 1.
The invention carries out the experiment of killing effect of detecting the bispecific antibody IgG-scFv on the tumor cells by lactate dehydrogenase, the experiment of promoting cytokine secretion of T cells by the bispecific antibody IgG-scFv, the experiment of inhibiting the tumor cell proliferation and the effect on a c-Met downstream signal path by the bispecific antibody IgG-scFv and the experiment of acting the bispecific antibody IgG-scFv in an NOD-SCID subcutaneous tumor-bearing mouse model, and the experiment shows that the bispecific antibody can recognize the c-Met on the surface of the tumor cells and block the combination of a negative co-stimulation molecule PD-1 on the surface of the T cells and PD-L1 on the surface of the tumor cells, so that the T cells generate continuous killing effect on the tumor cells.
Further, the present invention provides the use of said bispecific antibody against PD-1 antigen and against c-Met antigen in the manufacture of a medicament; the medicine is a tumor medicine for treating or preventing c-Met specific antigen expression.
On the basis of computer molecular structure simulation, the invention utilizes molecular cloning technology to construct expression plasmids of bispecific antibodies by taking two screened monoclonal antibodies as templates, wherein the expression plasmids are tetravalent bispecific antibodies, namely recombinant humanized PD-1 and c-Met bispecific antibodies, tests show that the expression plasmids can identify c-Met on the surface of tumor cells and block the combination of negative co-stimulation molecules PD-1 on the surface of T cells and PD-L1 on the surface of the tumor cells, so that the T cells generate continuous killing effect on the tumor cells, and the invention provides a basis for immune intervention of targeted c-Met positive tumor cells.
The following terms and abbreviations are used in the present invention:
PD-1: programmed cell death receptor 1(programmed cell death-1)
c-Met: hepatocyte growth factor receptor (mesenchymal transfer factor)
HGF: hepatocyte growth factor (hepatocyte growth factor)
IgG: immunoglobulin G (immunoglobulin Globulin G)
BsAb: bispecific antibody (bispecific antibody)
ScFv: single chain variable region antibody fragment (single-chain variable fragment)
VH: heavy chain variable region (heavy chain variable region)
VL: light chain variable region (light chain variable region)
EBNA 1: EB virus nuclear antigen (Epstein-Barr virus nuclear antigen 1)
HEK 293: human Embryonic Kidney 293 cell (Human Embryonic Kidney 293)
IFN-gamma: Gamma-Interferon (Interferon-gamma)
PEI: polyethyleneimine (Polyethylenimine)
FITC: fluorescein isothiocyanate (fluoroescein isothiocyanate)
PHA-L: phytohemagglutinin (Phytohemagglutinin-L)
SDS-PAGE: sodium dodecyl sulfate Polyacrylamide gel electrophoresis (sodium didecyl sulfate Polyacrylamide gel electrophoresis)
PI: propidium iodide (Propidium lodide)
PBS: phosphate buffered saline (phosphate buffer saline)
Drawings
FIG. 1 is a schematic diagram of the construction of a bispecific antibody against PD-1 antigen and c-Met antigen of the present invention.
FIG. 2 is a schematic structural diagram of an anti-PD-1 antigen and anti-c-Met antigen bispecific antibody of the present invention.
FIG. 3 is a SDS-PAGE electrophoretic analysis diagram of a bispecific antibody against PD-1 antigen and against c-Met antigen of the present invention.
FIG. 4 is a flow-binding diagram of a bispecific antibody against PD-1 antigen and against c-Met antigen of the present invention.
FIG. 5 is a graph showing the results of in vitro cytotoxicity analysis of a bispecific antibody against PD-1 antigen and against c-Met antigen of the present invention.
FIG. 6 is a graph showing the results of cytokine secretion analysis of a bispecific antibody against PD-1 antigen and against c-Met antigen of the present invention.
FIG. 7 is a graph showing the results of in vitro inhibition of tumor cell proliferation by a bispecific antibody against PD-1 antigen and against c-Met antigen of the present invention.
FIG. 8 is a graph showing the results of Western Blot analysis of the inhibition of the c-Met downstream signaling pathway by the bispecific antibody against PD-1 antigen and c-Met antigen of the present invention.
FIG. 9 is a graph comparing the inhibition of tumor proliferation in vivo by a bispecific antibody against PD-1 antigen and c-Met antigen of the present invention with PBS control and PBMC simplex.
Preferred embodiments of the present invention will be described in further detail below with reference to the accompanying drawings.
Detailed Description
Example 1: molecular design of bispecific antibody IgG-scFv and construction of expression plasmid
Through computer molecular structure simulation and sequence analysis, secretion signal peptide and vector expression element are optimized, expression plasmid of bispecific antibody IgG-scFv is designed, proper primer and overlap extension PCR are used to connect the variable regions of two antibodies separately, and heavy and light chains are constituted separately into eukaryotic expression vector pCEP4 through recombination reaction.
Example 2: expression of bispecific antibody IgG-scFv in host cell HEK293E cells
Cells were switched to serum-free culture 24hr before transfection, and the cell density was approximately 1X10 on the day of transfection6And (2) co-transfecting HEK293E cells with a heavy-light chain expression plasmid by using a cationic transfection reagent PEI at a ratio of 1: 1, adding 0.5% of tryptone Trypton N1 which is freshly prepared and subjected to filter sterilization after transfection is carried out for 24 hours, centrifuging to collect culture supernatant after the proportion of living cells is reduced to about 60%, and storing at-20 ℃ after filter sterilization for purification.
Example 3: protein A one-step purification of bispecific antibody IgG-scFv
A1 ml prepacked chromatography column of Protein A was connected to an AKTA Protein purification system, the column was equilibrated with PBS (pH7.4) and loaded, a flow-through sample was collected, the column was then washed with about 10 column volumes of PBS (pH7.4) to remove non-specifically bound contaminating proteins and a wash sample was collected, and finally the target Protein was eluted with 0.1M citric acid (pH3.0) and neutralized with 1M Tris-HCL (pH9.0), concentrated by ultrafiltration and stored by freeze-drying.
Example 4: bispecific antibody IgG-scFv antibody binding specificity flow assay
The purified bispecific antibody IgG-scFv was incubated with CHO stably expressing PD-1 extracellular domain, gastric cancer cell MKN45 expressing c-MET and isotype control IgG, respectively, and after FITC-labeled fluorescent secondary antibody was added, the fluorescence intensity was analyzed by flow cytometry.
Example 5: method for detecting killing effect of bispecific antibody IgG-scFv on tumor cells by lactate dehydrogenase
PHA-L stimulated hPBMCs were cultured 48hr before the experiment, MKN45 cells or MGC803 cells were plated in 96-well plates, PBMC was added at different ratios of effector cells to target cells, IgG-scFv diabodies were added, and controls were set according to the instructions. After mixing and incubation for 48hr, absorbance values were read at 490nm after addition of the corresponding reagent according to the kit instructions.
Example 6: bispecific antibody IgG-scFv function for promoting T cell to secrete cytokine
PBMC separated from human fresh blood samples and target cells (MKN45 or MGC803) are planted in a 96-well plate in different proportions, IgG-scFv double antibodies with different concentration gradients or IgG4 control with corresponding concentration are added, PHA-L with the final concentration of 1 mu g/ml is added after incubation for 30min, and cell culture supernatant is collected after standing and culturing for 72hr to detect the secretion levels of the cytokines IFN-gamma and Granzyme B according to ELISA kit instructions.
Example 7: CCK-8 detection of inhibition of bispecific antibody IgG-scFv on tumor cell proliferation
MKN45 cells were seeded in a 96-well plate, 0.5. mu.M or 1.0. mu.M IgG-scFv bispecific antibody and a control of the same concentration were added, and after standing and culturing for 12hr, CCK8 reagent was added, and after incubation for 4hr, absorbance was measured at 450nm with a microplate reader.
Example 8: western Blot for detecting effect of bispecific antibody IgG-scFv on c-Met downstream signal channel
Adjusting the cell density of MKN45 to 5x104, planting in a 6-well plate, changing into a serum-free culture medium after 24hr, adding IgG-scFv double antibody with a certain concentration gradient or a control with a corresponding concentration, incubating at 37 ℃ for 2hr, stimulating with HGF with a final concentration of 100ng.ml for 10min, lysing cells, centrifuging, collecting supernatant, performing SDS-PAGE electrophoresis, sealing with a transfer membrane, incubating with a corresponding phosphorylation or background antibody overnight, incubating with a corresponding secondary antibody for the next day, and observing the activation state of c-Met and downstream signal molecules thereof by chemiluminescence.
Example 9: role of bispecific antibody IgG-scFv in NOD-SCID subcutaneous tumor-bearing mouse model
Selecting 30 male NOD-SCID mice of 5-6 weeks, dividing into 2 groups, inoculating 5 × 10 in right axilla6The gastric cancer cells MKN45 and MGC803 are added until the tumor volume reaches about 50mm3Then, 15 mice in each group were randomly divided into 3 groups of 5 mice in each group, activated PBMCs were collected and centrifuged, and PBS was injected into tail vein of control group at 200 μ L/time; PBMC simplex group with 1X10 intravenous injections per mouse7PBMC, dual anti + PBMC experimental group: IgG-scFv diabody 100. mu.g, each mouse tail vein injected with 1X107PBMC were administered 2 times per week for 4 consecutive weeks during which the length a and width b of the subcutaneous tumor were measured with a vernier caliper and the tumor volume was calculated using the tumor volume formula, where the tumor volume was 0.5 × a × b2Tumor volume changes were recorded.
Claims (7)
1. A bispecific antibody against a PD-1 antigen and against a c-Met antigen, characterized in that it comprises: a first antibody segment specifically recognizing the PD-1 antigen of T lymphocytes and a second antibody segment specifically recognizing the c-Met antigen of tumor cells.
2. The bispecific antibody against PD-1 antigen and c-Met antigen according to claim 1, wherein the light chain unit of the bispecific antibody is the light chain variable region and the light chain constant region of the anti-PD-1 antibody, and the heavy chain unit of the bispecific antibody is: a C-Met scFv region is formed by a light chain variable region and a heavy chain variable region of the anti-C-Met antibody through a Gly4Ser connecting peptide, and the scFv region is connected to the C terminal of the heavy chain unit of the anti-PD-1 antibody through a Gly4Ser connecting peptide sequence to form the heavy chain unit of the bispecific antibody; wherein, the nucleotide sequence of the light chain unit of the anti-PD-1 antibody is shown as SEQ ID NO: 1, the nucleotide sequence of the heavy chain unit of the anti-PD-1 antibody is shown in SEQ ID NO: 2, the nucleotide sequence of the light chain variable region of the anti-c-Met antibody is shown as SEQ ID NO: 3, the nucleotide sequence of the heavy chain variable region of the anti-c-Met antibody is shown as SEQ ID NO: 4, and the nucleotide sequence of the Gly4Ser connecting peptide is shown as SEQ ID NO: 5, respectively.
3. The bispecific antibody against a PD-1 antigen and an anti c-Met antigen according to claim 1, characterized by being prepared by the following method:
(1) constructing the heavy chain and the light chain into the same expression vector respectively;
(2) co-transfecting the two expression vectors constructed in (1) together into a host cell, culturing and separating a supernatant;
(3) and purifying the separated supernatant to obtain the PD-1& c-Met bispecific antibody IgG-scFv.
4. The bispecific antibody against PD-1 antigen and c-Met antigen according to claim 3, wherein in the production method, the expression vector is pCEP 4.
5. The method of claim 3, wherein the host cell is a human embryonic kidney 293 cell HEK293E engineered with EBNA 1.
6. Use of the bispecific antibody against the PD-1 antigen and the c-Met antigen as claimed in any one of claims 1 to 4 in the manufacture of a medicament.
7. Use according to claim 6, wherein the medicament is a medicament for the treatment or prevention of tumours expressing c-Met specific antigens.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810748597.6A CN110684113A (en) | 2018-07-06 | 2018-07-06 | Bispecific antibody for resisting PD-1 antigen and c-Met antigen and construction method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810748597.6A CN110684113A (en) | 2018-07-06 | 2018-07-06 | Bispecific antibody for resisting PD-1 antigen and c-Met antigen and construction method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110684113A true CN110684113A (en) | 2020-01-14 |
Family
ID=69107012
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810748597.6A Pending CN110684113A (en) | 2018-07-06 | 2018-07-06 | Bispecific antibody for resisting PD-1 antigen and c-Met antigen and construction method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110684113A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114195900A (en) * | 2020-09-17 | 2022-03-18 | 普米斯生物技术(珠海)有限公司 | Anti-4-1 BB/PD-L1 bispecific antibody and application thereof |
| WO2022127842A1 (en) * | 2020-12-17 | 2022-06-23 | 上海华奥泰生物药业股份有限公司 | Bispecific antibody targeting il-17a and il-36r and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085680A (en) * | 2014-05-23 | 2015-11-25 | 复旦大学 | Humanized anti-PD-1 and c-MET bispecific antibody, and preparation method and application thereof |
| CN106967172A (en) * | 2016-08-23 | 2017-07-21 | 中山康方生物医药有限公司 | The anti-bifunctional antibodies of PD 1 of anti-CTLA 4, its medical composition and its use |
| CN107090038A (en) * | 2011-06-30 | 2017-08-25 | 霍夫曼-拉罗奇有限公司 | Anti- C MET antibody formulations |
| CN107847574A (en) * | 2015-07-30 | 2018-03-27 | 宏观基因有限公司 | The binding molecules of PD 1 and its application method |
-
2018
- 2018-07-06 CN CN201810748597.6A patent/CN110684113A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107090038A (en) * | 2011-06-30 | 2017-08-25 | 霍夫曼-拉罗奇有限公司 | Anti- C MET antibody formulations |
| CN105085680A (en) * | 2014-05-23 | 2015-11-25 | 复旦大学 | Humanized anti-PD-1 and c-MET bispecific antibody, and preparation method and application thereof |
| CN107847574A (en) * | 2015-07-30 | 2018-03-27 | 宏观基因有限公司 | The binding molecules of PD 1 and its application method |
| CN106967172A (en) * | 2016-08-23 | 2017-07-21 | 中山康方生物医药有限公司 | The anti-bifunctional antibodies of PD 1 of anti-CTLA 4, its medical composition and its use |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114195900A (en) * | 2020-09-17 | 2022-03-18 | 普米斯生物技术(珠海)有限公司 | Anti-4-1 BB/PD-L1 bispecific antibody and application thereof |
| CN114195900B (en) * | 2020-09-17 | 2024-02-23 | 普米斯生物技术(珠海)有限公司 | Anti-4-1 BB/PD-L1 bispecific antibody and application thereof |
| WO2022127842A1 (en) * | 2020-12-17 | 2022-06-23 | 上海华奥泰生物药业股份有限公司 | Bispecific antibody targeting il-17a and il-36r and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20220002418A1 (en) | Anti-pd-l1/vegf bifunctional antibody and use thereof | |
| US10118964B2 (en) | Construction and application of bispecific antibody HER2xCD3 | |
| US11286309B2 (en) | Antihuman PCSK9 monoclonal antibody and application thereof | |
| EA030147B1 (en) | Bispecific t cell activating antigen binding molecules | |
| CN104558191B (en) | Construction and application of bispecific antibody CD20 xCD 3 | |
| Han et al. | Recent clinical trials utilizing chimeric antigen receptor T cells therapies against solid tumors | |
| Wu et al. | HER2-specific chimeric antigen receptor-engineered natural killer cells combined with apatinib for the treatment of gastric cancer | |
| JP7461397B2 (en) | Immunomodulatory and antitumor related nanobodies and their nucleic acid coding sequences and uses thereof | |
| WO2023151346A1 (en) | Construction and application of novel bispecific chimeric antigen receptor | |
| CN110684113A (en) | Bispecific antibody for resisting PD-1 antigen and c-Met antigen and construction method thereof | |
| CN111944053B (en) | anti-BCMA CAR and expression vector and application thereof | |
| Yang et al. | Contextual reprogramming of CAR-T cells for treatment of HER2+ cancers | |
| CN110546165A (en) | anti-PD-L1-anti-TIM-3 bispecific antibody | |
| WO2020019983A1 (en) | Genetically engineered cell used for treating tumour | |
| CN102559636A (en) | Antibody fusion protein used for leukemia and autoimmune disease and preparation method thereof | |
| EP2091972B1 (en) | Multi-modal cancer therapy using viral hitch-hiking | |
| CN110564767A (en) | attenuated virus vector system, application of attenuated virus vector system in preparation of anti-malignant tumor medicine and use method of medicine | |
| EP4332116A1 (en) | Anti-cntn4-specific antibodies and use thereof | |
| US20230277668A1 (en) | Chimeric antigen receptor and use thereof | |
| CN111848806B (en) | EGFR-CD3 bifunctional antibody and application thereof | |
| US20230192898A1 (en) | Covalent multi-specific antibody | |
| CN115124625A (en) | Immune regulation and anti-tumor related nano antibody, nucleic acid coding sequence and application thereof | |
| WO2019157691A1 (en) | Recombinant chimeric antigen receptor gene and use thereof | |
| CN110734494A (en) | anti-TSPAN 8 monoclonal antibody and application thereof | |
| CN112239506B (en) | Small molecule bispecific antibody diabetes of anti-PD-1 and c-Met antigen |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200114 |