CN109420168A - A kind of biological agent technology of preparing inhibiting WT1 gene mutation - Google Patents
A kind of biological agent technology of preparing inhibiting WT1 gene mutation Download PDFInfo
- Publication number
- CN109420168A CN109420168A CN201710769228.0A CN201710769228A CN109420168A CN 109420168 A CN109420168 A CN 109420168A CN 201710769228 A CN201710769228 A CN 201710769228A CN 109420168 A CN109420168 A CN 109420168A
- Authority
- CN
- China
- Prior art keywords
- lymphocyte
- inhibiting
- cell
- biological agent
- gene mutation
- 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.)
- Withdrawn
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
-
- 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
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0636—T lymphocytes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/30—Coculture with; Conditioned medium produced by tumour cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Hematology (AREA)
- Mycology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Virology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
This application discloses a kind of preparation methods of biological agent for inhibiting WT1 gene mutation, comprising: T lymphocyte is isolated from blood of human body；T lymphocyte is placed in culture solution and carries out amplification cultivation；Cultured T lymphocyte is placed in culture bottle with cell strain and is tamed；Cell strain has been tamed T lymphocyte together with cancer cell to test, after T lymphocyte has the ability of attack cancer cell, has been separated；The T lymphocyte isolated and WT1 gene are subjected to hybrid test, the T lymphocyte after domestication has the ability for resisting WT1 gene mutation；These T lymphocytes for having resistance attacking ability are separated from culture solution again, active ingredient is made together with human immunoglobulin, is sealed refrigeration.The invention has the advantages that inhibiting the mutation of WT1 gene from source, the generation of cancer cell is prevented, effectively pre- anti-cancer, preventive effect can reach 95% or more, it is as a result more accurate.
Description
Technical field
The present invention relates to a kind of preparation methods of biological agent for inhibiting WT1 gene mutation.
Background technique
Currently, Cancer in China morbidity and mortality are rising always, have become one of China since 2010
Major public health problem.2 months 2017, National Cancer Center issued newest cancer data, summarized national 347 cancers
Register the data of point, data are shown: about 10,000 people make a definite diagnosis cancer daily in the whole nation；Lung cancer is disease incidence, the double rates first of the death rate；First
Shape gland cancer rapid increase.In the same year, world's new cases about 14,090,000, Chinese new cancer cases account for the 1/4 of the world.
Scientific research shows, various cancers all represented in fact with the mankind tumorigenic gene WT1 gene have it is direct
System, the cancer of either lung cancer, gastric cancer, thyroid cancer or other types are all related with this gene mutation.The prior art pair
There are many kinds of drugs for the prevention and treatment of cancer, but have no exact preventive effect for treating cancer, are mainly manifested in: first, it is right
The prevention effect of cancer is indefinite, can not effective pre- anti-cancer；Second, drug is very big to the destructive power of human immune system, portion
Divide patient that can not endure.
Summary of the invention
The purpose of the present invention is to overcome the above shortcomings and to provide a kind of preparations of biological agent for inhibiting WT1 gene mutation
Method can generate exact technical effect to pre- anti-cancer.
To achieve the goals above, a kind of the technical solution adopted by the present invention are as follows: biological agent for inhibiting WT1 gene mutation
Preparation method, which comprises the following steps: T lymphocyte is isolated from blood of human body；T lymphocyte is put
It is placed in culture solution and carries out amplification cultivation；Cultured T lymphocyte is placed in culture bottle with cell strain and is tamed；It will be thin
Born of the same parents' strain has been tamed T lymphocyte and has been tested together with cancer cell, after T lymphocyte has the ability of attack cancer cell, into
Row separation；The T lymphocyte isolated and WT1 gene are subjected to hybrid test, the T lymphocyte after domestication, which has, resists WT1
The ability of gene mutation；These T lymphocytes for having resistance attacking ability are separated from culture solution again, with human immunity
Active ingredient is made in globulin together, is sealed refrigeration.
The invention has the benefit that
Preparation method is simple, T lymphocyte is isolated from blood of human body, and carry out amplification cultivation, and domestication, separation mix
Active ingredient is made in the step of closing test, and human body is allowed to generate the memory t cell for inhibiting WT1 gene mutation.It is controlled from source
WT1 gene does not allow it to be mutated, prevents the generation of cancer cell, effectively pre- anti-cancer, preventive effect can reach 95% or more, knot
Fruit is more accurate.
Specific embodiment
Specification subsequent descriptions are to implement the better embodiment of the application, and so the description is one to illustrate the application
As for the purpose of principle, be not intended to limit the scope of the present application.The protection scope of the application is worked as to be defined depending on appended claims
Subject to person.
The preparation method of the biological agent of inhibition WT1 gene mutation of the invention, comprising the following steps: from blood of human body
Isolate T lymphocyte；T lymphocyte is placed in culture solution and carries out amplification cultivation；By cultured T lymphocyte and carefully
Born of the same parents' strain, which is placed in culture bottle, tames；Cell strain is tamed T lymphocyte together with cancer cell to test, when T lymph
After cell has the ability of attack cancer cell, separated；The T lymphocyte isolated is carried out to mix survey with WT1 gene
Examination, the T lymphocyte after domestication have the ability for resisting WT1 gene mutation；These are had to the T leaching for resisting attacking ability again
Bar cell is separated from culture solution, and active ingredient is made together with human immunoglobulin, is sealed refrigeration.
Described the step of T lymphocyte is isolated from blood of human body includes hundred grades of lifes in ten thousand grades of bio clean rooms of GMP
The operation of object clean bench, operation temperature is room temperature.
Preferably, described the step of T lymphocyte is placed in progress amplification cultivation in culture solution includes: in KRA cell
Amplification cultivation, the KRA cell culture fluid ingredient percent are carried out in culture solution are as follows:
Preferably, the environmental condition of the amplification cultivation is 37 degree of amplification cultivations of carbon dioxide incubator temperature.
Preferably, described that cultured T lymphocyte and cell strain be placed in culture bottle to carry out domestication include: so that T
Lymphocytic cell surface forms membrane molecule, participates in cell recognition antigen, cell activation, proliferation, differentiation；And use back-and-forth method or clone
Formation method obtains the cell strain with special nature or marker from primary culture or cell line.
Preferably, the environment of the domestication is nontoxic, sterile, and temperature is 37 degree.
Preferably, described that cell strain has been tamed into the step of T lymphocyte is tested together with cancer cell, the test
Condition be to be carried out under 37 degree of gnotobasis, and separated using immunomagnetic beads mode.
Preferably, the step of T lymphocyte isolated and WT1 gene being subjected to hybrid test, the hybrid test
Condition is carried out under 37 degree of gnotobasis.
Preferably, the formula of the mixed solution is selected from the one or more of following component: calcium chloride, calcium nitrate, sulfuric acid
Magnesium, five H 2 O calcium sulphates, ferrous sulfate heptahydrate, white vitriol, L-alanine, L- days (door) winters amide-hydrate, L- asparagus fern
Propylhomoserin, L-cysteine-hydrochloride monohydrate, l-cysteine-dihydrochloride, Pidolidone, glycine, L-Histidine-salt
Hydrochlorate-hydrate, L- hydroxyproline, l-Isoleucine, L-Leu, L-lysine-hydrochloride, L-Methionine, Agifutol
Reduced form, ascorbic acid, biotin, vitamin B12, D-VB5 salt calcium, putrescine (putrescine)-dihydrochloride, chlorination
Choline, folic acid, i-Inositol, niacinamide, pyridoxal-hydrochloride, vitamin B6-hydrochloride, vitamin B2, thiamines-hydrochloric acid
Salt, lipoic acid, Para-Aminobenzoic, adenosine, uridine, cytidine, guanosine.
Preferably, the condition of the culture bottle is that have good transparency and nontoxic, sterile outer, is also needed surface-modified
Processing enables adherent, division and growth.
Preferably, the T lymphocyte that these are had resistance attacking ability is separated from culture solution, with human immunity
It is room temperature centrifugation, the preparation condition of active ingredient that the condition that the step of active ingredient includes: separation, which is made, in globulin together
It is hundred grades of biologic cleanliness platforms operation in ten thousand grades of bio clean rooms of GMP, operation temperature is room temperature.
Test effect:
The oral medicinal herb of biological agent and the prior art of the invention is prevented into cancer group and Western medicine preparation prevents cancer group
It is compared as follows:
Above-mentioned preparation method is simple, T lymphocyte is isolated from blood of human body, and carry out amplification cultivation, and domestication divides
Active ingredient is made in the step of from, hybrid test, and human body is allowed to generate the memory t cell for inhibiting WT1 gene mutation.From source
Control WT1 gene, do not allow it be mutated, prevent the generation of cancer cell, can effectively pre- anti-cancer, preventive effect reach 95% with
On, it is as a result more accurate.
Explanation shows and describes several preferred embodiments of the present application, but as previously described, it should be understood that the application is not
It is confined to form disclosed herein, should not be regarded as an exclusion of other examples, and can be used for various other combinations, modification
And environment, and can be carried out in application contemplated scope described herein by the above teachings or related fields of technology or knowledge
Change.And changes and modifications made by those skilled in the art do not depart from spirit and scope, then it all should be in the application institute
In attached scope of protection of the claims.
Claims (9)
1. a kind of preparation method for the biological agent for inhibiting WT1 gene mutation, which comprises the following steps:
T lymphocyte is isolated from blood of human body；
T lymphocyte is placed in culture solution and carries out amplification cultivation；
Cultured T lymphocyte is placed in culture bottle with cell strain and is tamed；
Cell strain is tamed T lymphocyte together with cancer cell to test, when T lymphocyte has the energy of attack cancer cell
After power, separated；
The T lymphocyte isolated and WT1 gene are subjected to hybrid test, the T lymphocyte after domestication, which has, resists WT1 gene
The ability of mutation；
These T lymphocytes for having resistance attacking ability are separated from culture solution again, together with human immunoglobulin
Active ingredient is made, is sealed refrigeration.
2. the preparation method of the biological agent according to claim 1 for inhibiting WT1 gene mutation, which is characterized in that described
The step of T lymphocyte is isolated from blood of human body includes hundred grades of biologic cleanliness platforms operation in ten thousand grades of bio clean rooms of 6MP,
Operation temperature is room temperature.
3. the preparation method of the biological agent according to claim 2 for inhibiting WT1 gene mutation, which is characterized in that described
The step of T lymphocyte is placed in progress amplification cultivation in culture solution includes: that amplification training is carried out in KRA cell culture fluid
It supports, the KRA cell culture fluid ingredient percent are as follows:
4. the preparation method of the biological agent according to claim 3 for inhibiting WT1 gene mutation, which is characterized in that described
The environmental condition of amplification cultivation is 37 degree of amplification cultivations of carbon dioxide incubator temperature.
5. the preparation method of the biological agent according to claim 1 for inhibiting WT1 gene mutation, which is characterized in that described
It includes: so that T lymphocyte surface forms film point that cultured T lymphocyte and cell strain, which are placed in culture bottle, and carry out domestication
Son participates in cell recognition antigen, cell activation, proliferation, differentiation；And using back-and-forth method or clone forming method from primary culture or
The cell strain with special nature or marker is obtained in cell line.
6. the preparation method of the biological agent according to claim 5 for inhibiting WT1 gene mutation, which is characterized in that described
The environment of domestication be it is nontoxic, sterile, temperature be 37 degree.
7. the preparation method of the biological agent according to claim 6 for inhibiting WT1 gene mutation, which is characterized in that described
Cell strain the step of T lymphocyte is tested together with cancer cell is tamed into, the condition of the test is sterile at 37 degree
It carries out under environment, and is separated using immunomagnetic beads mode.
8. the preparation method of the biological agent according to claim 6 for inhibiting WT1 gene mutation, which is characterized in that will divide
The step of T lymphocyte and WT1 gene separated out carries out hybrid test, the condition of the hybrid test is in 37 degree of gnotobasis
Lower progress.
9. the preparation method of the biological agent according to claim 3 for inhibiting WT1 gene mutation, which is characterized in that described
The formula of mixed solution is selected from the one or more of following component:
Calcium chloride, calcium nitrate, magnesium sulfate, five H 2 O calcium sulphates, ferrous sulfate heptahydrate, white vitriol, L- alanine, L- days (door)
Winter amide-hydrate, L-Aspartic acid, L-cysteine-hydrochloride monohydrate, l-cysteine-dihydrochloride, Pidolidone,
Glycine, L-Histidine-hydrochloride monohydrate, L- hydroxyproline, l-Isoleucine, L-Leu, L-lysine-hydrochloride,
L-Methionine, Agifutol reduced form, ascorbic acid, biotin, vitamin B12, D-VB5 salt calcium, putrescine are (rotten
Amine)-dihydrochloride, choline chloride, folic acid, i-Inositol, niacinamide, pyridoxal-hydrochloride, vitamin B6-hydrochloride, dimension
Raw element B2, thiamines-hydrochloride, lipoic acid, Para-Aminobenzoic, adenosine, uridine, cytidine, guanosine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710769228.0A CN109420168A (en) | 2017-08-31 | 2017-08-31 | A kind of biological agent technology of preparing inhibiting WT1 gene mutation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710769228.0A CN109420168A (en) | 2017-08-31 | 2017-08-31 | A kind of biological agent technology of preparing inhibiting WT1 gene mutation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109420168A true CN109420168A (en) | 2019-03-05 |
Family
ID=65504679
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710769228.0A Withdrawn CN109420168A (en) | 2017-08-31 | 2017-08-31 | A kind of biological agent technology of preparing inhibiting WT1 gene mutation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109420168A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040161433A1 (en) * | 1999-10-21 | 2004-08-19 | Keisuke Teshigawara | Method for in vitro culture of lymphocytes and composition for use in immune therapy |
| US20150344844A1 (en) * | 2014-02-04 | 2015-12-03 | Marc Better | Methods for producing autologous t cells useful to treat b cell malignancies and other cancers and compositions thereof |
| US20170042995A1 (en) * | 2014-04-30 | 2017-02-16 | President And Fellows Of Harvard College | Combination Vaccine Devices and Methods of Killing Cancer Cells |
| US20170137783A1 (en) * | 2015-07-21 | 2017-05-18 | Felipe Bedoya | Methods for improving the efficacy and expansion of immune cells |
-
2017
- 2017-08-31 CN CN201710769228.0A patent/CN109420168A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040161433A1 (en) * | 1999-10-21 | 2004-08-19 | Keisuke Teshigawara | Method for in vitro culture of lymphocytes and composition for use in immune therapy |
| US20150344844A1 (en) * | 2014-02-04 | 2015-12-03 | Marc Better | Methods for producing autologous t cells useful to treat b cell malignancies and other cancers and compositions thereof |
| US20170042995A1 (en) * | 2014-04-30 | 2017-02-16 | President And Fellows Of Harvard College | Combination Vaccine Devices and Methods of Killing Cancer Cells |
| US20170137783A1 (en) * | 2015-07-21 | 2017-05-18 | Felipe Bedoya | Methods for improving the efficacy and expansion of immune cells |
Non-Patent Citations (3)
| Title |
|---|
| 杨吉成等: "《医用细胞工程》", 31 December 2003, 上海交通大学出版社 * |
| 顾伟英等: "WT1基因与恶性肿瘤靶向免疫治疗", 《肿瘤》 * |
| 顾伟英等: "WT1基因肽诱导细胞毒性T淋巴细胞免疫治疗白血病的实验研究", 《中华医学杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TWI797067B (en) | Compositions comprising bacterial strains | |
| Kale et al. | The diatoms: Big significance of tiny glass houses | |
| CN112175864B (en) | Bifidobacterium animalis and breeding method and application thereof | |
| CN105039253B (en) | Serum free medium suitable for immunocyte large-scale culture | |
| CN104224851B (en) | The application of bifidobacterium animalis acid subspecies CGMCC No.9273 | |
| Rees | The antibody molecule: from antitoxins to therapeutic antibodies | |
| CN105624069A (en) | Lactobacillus rhamnosus preparation and application thereof | |
| CN107208044A (en) | It is engineered to treat the bacterium of the disease related to hyperammonemia | |
| Paul et al. | Antibiotics associated disorders and post-biotics induced rescue in gut health | |
| CN109420168A (en) | A kind of biological agent technology of preparing inhibiting WT1 gene mutation | |
| CN104940450A (en) | Health beverage for enhancing body immunity and preparation method thereof | |
| SE526711C2 (en) | Novel strains of Bifidobacterium having ability to survive in intestinal tract and produce glutamine and arginine in vivo, useful for preparing medicament for treatment of intensive care patients with intestinal failure | |
| Taoka et al. | Production of anserine-rich fish sauce from giant masu salmon, Oncorhynchus masou masou and γ-Aminobutyric Acid (GABA)-enrichment by Lactobacillus plantarum Strain N10 | |
| CN110269934A (en) | A kind of preparation method of the biological agent of WT1 gene anticancrin | |
| Davis et al. | Immune status of patients with multiple sclerosis: Analysis of primary and established immune responses in 24 patients | |
| CN106389475B (en) | Bacteroides fragilis is preventing and/or is treating the application in meningitis | |
| CN106591177A (en) | Mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium and preparation method thereof | |
| Murray et al. | Possible role of the microbiome in the development of acute malnutrition and implications for food-based strategies to prevent and treat acute malnutrition | |
| CN113423413A (en) | Compositions comprising bacterial strains | |
| RU2657819C1 (en) | Means of immunomodulating activity | |
| Zeng et al. | The effect of artificial rearing on gut microbiota in a mouse pup-in-a-cup model | |
| Momot et al. | Practical use of the vaccination indicator in poultry farming | |
| CN107495335A (en) | One kind chelating calcium composition and preparation method thereof | |
| CN105968194B (en) | A kind of animal multiple-effect polyclonal antibody, preparation method and its application | |
| RU2409376C2 (en) | General health-improving agent in treating new growths |
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 | ||
| DD01 | Delivery of document by public notice | ||
| DD01 | Delivery of document by public notice |
Addressee: Liu Yichen Document name: Notification of Passing Examination on Formalities |
|
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190305 |