CN105838680B - One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application - Google Patents
One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application Download PDFInfo
- Publication number
- CN105838680B CN105838680B CN201610324645.XA CN201610324645A CN105838680B CN 105838680 B CN105838680 B CN 105838680B CN 201610324645 A CN201610324645 A CN 201610324645A CN 105838680 B CN105838680 B CN 105838680B
- Authority
- CN
- China
- Prior art keywords
- estriol
- monoclonal antibody
- cell strain
- antibody specific
- hybridoma cell
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/44—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
Abstract
One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application, belong to food safety technical field of immunoassay.Anti- estriol monoclonal antibody specific hybridoma cell strain NaN-2 of the invention, has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.12016.Anti- estriol monoclonal antibody specific, it is secreted by the hybridoma cell strain NaN-2 generates.The monoclonal antibody of this cell strain secretion has preferable specificity (IC to estriol50Value is 0.25ng/mL), it can be used for the specific detection of estriol in food safety.The anti-estriol cell strain of monoclonal antibody that the present invention obtains, there are preferable detection sensitivity and affinity to estriol, additionally provide a kind of method of new synthesis estriol immunogene, the synthesis step of haptens more simplifies, effectively, the thinking and method of synthetic immunogen are provided for the research of people from now on.
Description
Technical field
The present invention relates to one plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its applications, belong to
Food safety technical field of immunoassay.
Background technique
Estriol (E3) it is a kind of Endogenous steroids, it is estradiol (E2), oestrone (E1) metabolin, in pregnant woman's body
E3It is mainly derived from placenta, it is the important Testing index of late pregnancy development of fetus situation and placental function, and makeup
The common index that estrogen detects in product.Estriol be used to promote domestic animal growth and development in animal husbandry, and the improper of estriol makes
With the estriol residual contamination that may result in aquatic products, meat, eggs and newborn class.Clearly forbid at present edible in China
Estriol is added in the raising of animal.
Estriol detection at present mostly uses high performance liquid chromatography, liquid chromatogram and Mass Spectrometry, gas chromatography mass spectrometry method, puts
Penetrate immunization, enzyme-linked immunization, Chemiluminescence immunoassay, capillary electrophoresis etc..It is domestic generally to use GB/T 21981-2008
" hormone residues detection method in animal-derived food: one mass spectrum of liquid chromatogram, one mass spectrography " is detected, and homogeneous, enzyme need to be passed through
Solution, extraction, solid phase extraction concentration purification, Instrument measuring, interior scalar quantity.Clinically mainly detected using radioimmunology
Estriol, this method the disadvantages of there are radioactive isotope waste disposal problem and short marker storage lives.Above-mentioned detection method
Can carry out quantitative analysis and there is lower detection limit, but usually require the operation of expensive instrument and complexity, pre-treatment and
Detection time is long, seriously constrains the popularization of these detection methods.And immunoassay method has low cost, high-throughput, Gao Ling
The features such as quick, low to technical staff's relative requirement, therefore it is suitable for the rapid screening of a large amount of samples.It is an object of the invention to mention
There is the preparation method of the monoclonal antibody hybridoma cell strain of higher affinity and detection sensitivity for a kind of pair of estriol.For
The research and development popularization of indirect competitive ELISA kit and colloidal gold strip is laid a good foundation.
Summary of the invention
The object of the present invention is to provide one plant of anti-estriol monoclonal antibody specific hybridoma cell strains, by the cell strain
The antibody of preparation has preferable affinity and detection sensitivity to estriol, can be used to establish estriol enzyme linked immunosorbent detection
Method, or establish colloidal gold immunochromatographimethod technology rapid detection method.
Technical solution of the present invention, one plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2, preservation
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, abbreviation CGMCC, deposit number CGMCC
No.12016。
Anti- estriol monoclonal antibody specific, it is special by the anti-estriol that the deposit number is CGMCC No.12016
Specific monoclonal antibodies hybridoma cell strain NaN-2 secretion generates.
The application of the anti-estriol monoclonal antibody specific: its answering in estriol residue detection in food safety
With.
The preparation basic step of NaN-2 cell strain provided by the invention are as follows:
(1) preparation with identification of immunogene: estriol reacts preparation haptens containing carboxyl with 6- bromocaproic acid, passes through carbonization two
Imines method is connected with the amino of protein carrier, after reaction, passes through dialysis separation comlete antigen and the small molecule not being coupled half
Antigen, comlete antigen are identified by UV absorption scan method；
(2) mouse is immune: after immunogene and freund adjuvant emulsification completely, BALB/c is immunized by subcutaneous multi-point injection
Mouse.First immunisation uses Freund's complete adjuvant, and booster immunization uses freund 's incomplete adjuvant, and immunizing dose is when spurt is immune
The half of a preceding immunizing dose is directly injected intraperitoneally after mixing with physiological saline；Each secondary immunization interval is three weeks.
After third time is immune, interval blood sampling in one week detection serum titer and inhibition；
(3) cell fusion and cell strain are established: by polyethylene glycol (PEG 4000) method, making mouse boosting cell and Mouse Bone
Myeloma cells fusion detects positive cell hole using indirect ELISA by HAT culture medium culture, and further using indirectly competing
Strive ELISA method measurement positive cell hole inhibitory effect, by limiting dilution assay to have the positive cell hole preferably inhibited progress
It is subcloned three times, finally screens and obtain hybridoma cell strain NaN-2；
(4) it the identification of hybridoma cell strain property: is set with and is surveyed with ELIAS secondary antibody using mouse monoclonal Ig class/subgroup identification
It is fixed；IC50The measurement of value, cross reacting rate and affinity passes through ELISA method.
Beneficial effects of the present invention: the anti-estriol cell strain of monoclonal antibody that the present invention obtains has preferably estriol
Detection sensitivity and affinity, additionally provide a kind of new method of synthesis estriol immunogene, the synthesis step of haptens
More simplify, effectively, provides the thinking and method of synthetic immunogen for the research of people from now on.
Biological material specimens preservation: monoclonal cell strain NaN-2 has been preserved in Chinese microorganism strain preservation conservator
Meeting common micro-organisms center, abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences microorganism
Research institute, deposit number are CGMCC No.12016, preservation date on January 20th, 2016.
Detailed description of the invention
The ultra-violet absorption spectrum of Fig. 1 immunogene characterizes.
Standard suppression curve of Fig. 2 NaN-2 monoclonal antibody to estriol.
Specific embodiment
The following examples of the present invention are only used as the further explanation of the content of present invention, not as a limitation of the present invention interior
Perhaps range.Below by embodiment, the invention will be further described.
The present invention is by being immunized mouse for estriol comlete antigen, by cell fusion, HAT selective medium culture,
Cell conditioned medium is screened by indirect ELISA and indirect competitive ELISA, having finally obtained has preferable affinity and detection to estriol
The monoclonal antibody hybridoma cell strain of sensitivity.
The preparation of 1 hybridoma cell strain NaN-2 of embodiment
(1) synthesis of comlete antigen: 300mg(1.10 mmol) E3It is dissolved in the dry dimethyl sulfoxide of 6mL, 1g is added
KOH powder.300mg bromocaproic acid is added after stirring 5min.Continue to stir, 50mL ice water is added after reacting 2h.Ethyl acetate extraction
Recycle unreacted E3.Water phase is acidified with 2mol/L HCl, white precipitate occurs.Sand core funnel filtering, sediment distilled water
It is washed till neutrality, is dried in vacuo.Methanol-chloroform recrystallization, obtains clear crystal, as estriol haptens.Take 4.5mg above-mentioned half
Antigen adds 2.0mg EDC(1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride) and 1.0mg NHS(N- hydroxyl
Base succinimide), use DMF(N, dinethylformamide) dissolution, it is stirred at room temperature, activates 4h；Separately take 5mg BSA(ox blood
Pure albumen) it is dissolved in the CB(carbonate buffer solution of 2mL, 0.05M, pH9.6) in solution, above-mentioned activating solution is added dropwise
It in BSA solution, is stirred at room temperature after reaction overnight, 4 DEG C are dialysed three days, and -20 DEG C of packing save.
(2) animal immune: the BALB/c mouse of 6~8 week old of health is selected to be immunized.Take estriol comlete antigen
After the emulsification uniformly of (1mg/mL) and equivalent freund adjuvant, BALB/c mouse, every 100 μ L are immunized by subcutaneous multi-point injection.It is first
It is secondary it is immune use Freund's complete adjuvant, booster immunization uses freund 's incomplete adjuvant, and immunizing dose is preceding primary when spurt is immune
The half of immunizing dose is directly injected intraperitoneally after mixing with physiological saline；Each secondary immunization interval is three weeks.For the third time
After immune, interval blood sampling in one week detection serum titer and inhibition；Selection inhibits best mouse, and spurt in 21 days is exempted from after exempting from five
Epidemic disease prepares fusion.
(3) cell fusion: after spurt immune three days, according to conventional PEG(polyethylene glycol, molecular weight 4000) method into
Row cell fusion, the specific steps are as follows:
A, sterile to take mouse spleen, it grinds and obtains splenocyte suspension by 200 mesh cell screen clothes, and carry out cytometer
Number；
B, SP2/0 cell is collected, is suspended in RPMI-1640 basic culture solution, cell count is carried out；
C, splenocyte and SP2/0 cell are mixed according to the ratio counted than 5-10:1, is merged after centrifugation with PEG, the time
RPMI-1640 basic culture solution is added later according to from slowly to fast in 1min, is suspended in after centrifugation containing 20% fetal calf serum, 2%
In the RPMI-1640 screening and culturing liquid of 50 × HAT, 96 porocyte culture plates are added to, 37 DEG C, 5%CO are placed in2Incubator in train
It supports.
(4) cell screening and cell strain are established: carrying out RPMI-1640 screening to fused cell in the third day of cell fusion
Culture solution partly changes liquid, carries out within the 5th day being carried out entirely with the RPMI-1640 transition culture solution of 100 × HT containing 20% fetal calf serum, 1%
Liquid is changed, took cell conditioned medium to be screened at the 7th day.Screen in two steps: the first step first filters out positive cell with indirect ELISA
Hole, it is standard items that second step, which selects estriol, carries out inhibitory effect measurement to positive cell hole with indirect competitive ELISA.Selection
There is the cell hole preferably inhibited to estriol, be subcloned using limiting dilution assay, is detected with same method.It repeats
Three times, cell strain NaN-2 is obtained.
(5) preparation and identification of monoclonal antibody: taking 8-10 week old BALB/c mouse, and every mouse peritoneal injects paraffin oil
1mL；Every mouse peritoneal injection 1 × 10 after 7 days6Hybridoma collected ascites since the 7th day, ascites was passed through pungent
Acid-saturated ammonium sulfate method purifying, the monoclonal antibody of acquisition are placed in -20 DEG C of preservations.
It is sub- that immunoglobulin is carried out using the monoclonal antibody that mouse monoclonal subtype identification kit obtains ascites purifying
Type identification, hypotype are IgG1 type.
Using indirect competitive ELISA method, monoclonal antibody is measured to the IC of estriol50For 0.25 μ g/L, it can be used for female three
The quickly detection of the specificity of alcohol.
(6) antibody application
Hybridoma cell strain NaN-2 is added by monoclonal antibody prepared by internal ascites applied to estriol ELISA
Recovery test, the specific steps are as follows:
The 6.1 0.1 μ g/mL for using carbonate buffer solution (CBS) to dilute as coating 96 hole elisa Plates of primordial covering, every hole
After 100 μ L, 37 DEG C of 2 h of coating, three times with PBST washing lotion board-washing, each every 250 μ L of hole, 3 min, is patted dry every time；
6.2 are closed with the CBS containing 0.2% gelatin, every hole 200 μ L, 37 DEG C of 2 h of closing, with PBST washing lotion board-washing three
Secondary, each every 250 μ L of hole, 3 min, pats dry every time；
6.3 0,0.2,0.5,1,2,5,10,20ng/mL estriol standard is respectively configured with phosphate buffer (PBS)
Solution.Standard solution and sample to be tested extracting solution are added separately in the ELISA Plate closed, every 50 μ L of hole,
Each sample repeats 3 holes, then the diluted anti-estriol monoclonal antibody of 50 μ L, 1 ︰ 16000,37 DEG C of reactions 0.5 are added in every hole
After h, board-washing is patted dry；
The sheep anti-mouse igg two that 100 μ L use the diluted HRP label of 1 ︰ of PBS 3000 containing 0.1% gelatin is added in 6.4 every holes
Anti-, after 37 DEG C of 0.5 h of reaction, board-washing is patted dry；
6.5 every holes are added 100 μ L TMB developing solutions, after 37 DEG C of 15 min of colour developing, 50 μ L 2M H of every hole addition2SO4It terminates
Liquid, 450 nm survey light absorption value；
6.6 addition recycling and sample pre-treatments: weigh 1g milk sample merging 50mL centrifuge tube in, respectively add 0.2ng,
0.4 ng and 1ng estriol.After the concussion uniformly of 25 mL, 80% methanol PBS solution (w/v) is added to sample, ultrasonic extraction 15
Min, 0.45 μm of membrane filtration of supernatant after standing, after filtrate uses the PBS containing 0.01% gelatin to dilute 4 times, as ELISA sample
Extracting solution is added recovery test using indirect competitive ELISA, and the rate of recovery is respectively 83%, 87%, 84%.
The configuration of solution:
Carbonate buffer solution (CBS): Na is weighed2CO31.59g NaHCO32.93g is mixed after being dissolved in a small amount of distilled water respectively
It closes, adds distilled water to mix to about 800mL, adjust pH value to 9.6, add distilled water to be settled to 1000mL, 4 DEG C of storages are spare；
Phosphate buffer (PBS): 8.00 g NaCl, 0.2 g KCl, 0.24 g KH2PO4, 3.62 g Na2HPO4·
12 H2O is dissolved in 800 mL pure water, with NaOH or HCl tune pH to 7.2~7.4, is settled to 1000 mL；
PBST: the PBS containing 0.05% Tween 20；
TMB developing solution: A liquid: Na2HPO4·12H2O 18.43g, citric acid 9.33g, pure water are settled to 1000 mL；B
Liquid: 60 mg TMB are dissolved in 100 mL ethylene glycol.A, B liquid 1 ︰ 5 mixing by volume is TMB developing solution, current existing mixed.
The subtype identification of 1. NaN-2 monoclonal antibody of table
2. NaN-2 monoclonal antibody of table is to oestrone, the IC of estradiol50And cross reacting rate
It is in summary only presently preferred embodiments of the present invention, practical range not for the purpose of limiting the invention.It is i.e. all
Equivalent changes and modifications made by content according to scope of the present invention patent all should be technology scope of the invention.
Claims (3)
1. one plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 has been preserved in Chinese microorganism strain guarantor
Administration committee's common micro-organisms center, abbreviation CGMCC are hidden, deposit number is CGMCC No.12016.
2. anti-estriol monoclonal antibody specific, it is characterised in that: it is CGMCC No.12016's by the deposit number
Anti- estriol monoclonal antibody specific hybridoma cell strain NaN-2 secretion generates, and has preferable specificity to estriol,
IC50Value is 0.25ng/mL.
3. the application of anti-estriol monoclonal antibody specific described in claim 2, it is characterised in that: it is female in food safety
Application in triol residue detection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610324645.XA CN105838680B (en) | 2016-05-17 | 2016-05-17 | One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610324645.XA CN105838680B (en) | 2016-05-17 | 2016-05-17 | One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105838680A CN105838680A (en) | 2016-08-10 |
| CN105838680B true CN105838680B (en) | 2019-05-24 |
Family
ID=56592678
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610324645.XA Active CN105838680B (en) | 2016-05-17 | 2016-05-17 | One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105838680B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106680516B (en) * | 2016-12-05 | 2018-07-06 | 北京勤邦生物技术有限公司 | Detect enzyme linked immunological kit and its application of estriol |
| CN116836280B (en) * | 2023-09-01 | 2023-10-31 | 北京纳百生物科技有限公司 | Anti-estriol monoclonal antibody, detection reagent and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010009514A1 (en) * | 2008-07-25 | 2010-01-28 | Newcastle Innovation Limited | Methods and kits for predicting the onset of labour |
| CN102426252A (en) * | 2011-08-31 | 2012-04-25 | 内蒙古科慧生物科技有限责任公司 | Kit for quantitative measurement of free estriol (FE3) and its detection method |
-
2016
- 2016-05-17 CN CN201610324645.XA patent/CN105838680B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010009514A1 (en) * | 2008-07-25 | 2010-01-28 | Newcastle Innovation Limited | Methods and kits for predicting the onset of labour |
| CN102426252A (en) * | 2011-08-31 | 2012-04-25 | 内蒙古科慧生物科技有限责任公司 | Kit for quantitative measurement of free estriol (FE3) and its detection method |
Non-Patent Citations (2)
| Title |
|---|
| 抗雌三醇单克隆抗体的研制;秦鸿志等;《生殖与避孕》;19891231;第9卷(第1期);第57-59页 * |
| 雌三醇单克隆抗体的制备与表征;王永成等;《北京大学学报》;20020731;第38卷(第4期);第459-465页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105838680A (en) | 2016-08-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105838681B (en) | One plant of anti-dexamethasone monoclonal antibody specific hybridoma cell strain C3 and its application | |
| CN107022527A (en) | Hybridoma, c reactive protein detection reagent of anti-c reactive protein monoclonal antibody and its preparation method and application can be secreted | |
| CN106226522A (en) | The detection method of a kind of GP73, detectable and detection kit | |
| CN113637081A (en) | Hybridoma cell strain secreting pendimethalin-resistant monoclonal antibody and application thereof | |
| CN104312978B (en) | A kind of TOB monoclonal antibody and preparation method and application | |
| CN105838680B (en) | One plant of anti-estriol monoclonal antibody specific hybridoma cell strain NaN-2 and its application | |
| US20220010029A1 (en) | Hybridoma cell strain that secrets anti-dinitolmide monoclonal antibodies and the application of hybridoma cell strain | |
| KR102189893B1 (en) | Antibody specifically binding a bPAG1 and use thereof | |
| CN112574956B (en) | Hybridoma cell strain secreting propamocarb monoclonal antibody and application thereof | |
| CN110343669B (en) | Hybridoma cell strain DNC secreting anti-triclabendazole monoclonal antibody and application thereof | |
| CN105907725B (en) | One plant of anti-hydrocortisone monoclonal antibody specific hybridoma cell strain YH7 and its application | |
| CN109705220A (en) | One plant of hybridoma cell strain for secreting anti-chlorine promazine monoclonal antibody and its application | |
| CN115340986B (en) | Hybridoma cell strain secreting monoclonal antibody of phorate and application thereof | |
| CN114752568B (en) | Furosemide monoclonal antibody, hybridoma cell strain and application | |
| CN114058594B (en) | Hybridoma cell strain secreting vitamin A monoclonal antibody and application thereof | |
| CN106929477B (en) | Anti-prostaglandin F2αSpecific monoclonal antibody hybridoma cell strain WXX-2 and application thereof | |
| CN104004718A (en) | Universal monoclonal antibody hybridoma cell strain capable of resisting pirlimycin and application thereof | |
| CN113637642A (en) | Hybridoma cell strain capable of secreting monoclonal antibody of dicofol and application of hybridoma cell strain | |
| FI96433B (en) | Monoclonal antibodies for the selective immunological determination of intact procollagen peptide (type III) and procollagen (type III) in body fluids | |
| JP4663831B2 (en) | Monoclonal antibodies, cell lines, and methods for measuring N1, N12-diacetylspermine | |
| CN108132348B (en) | Sudan red III hapten, coupling antigen, antibody and colloidal gold rapid detection device and application thereof | |
| CN110927376A (en) | Magnetic immunochemiluminescence detection kit for olaquindox and application thereof | |
| CN111454912A (en) | Cyperazine monoclonal antibody hybridoma cell strain and application thereof | |
| CN113754568B (en) | Acid orange I hapten, artificial antigen, synthesis and application thereof | |
| CN111217910A (en) | Monoclonal antibody pair and application thereof in detecting myeloperoxidase protein |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210425 Address after: 518000 3 / F, building 30, Baotian Industrial Zone, 8 Baotian lane, chentian community, Xixiang street, Bao'an District, Shenzhen City, Guangdong Province Patentee after: Shenzhen heavy chain Biotechnology Co.,Ltd. Address before: Food College of Jiangnan University No. 1800 Li Lake Avenue 214122 in Jiangsu province Wuxi City Binhu District Patentee before: Jiangnan University |
|
| TR01 | Transfer of patent right |