CN101614737A - A kind of colloidal gold strip and its production and application - Google Patents
A kind of colloidal gold strip and its production and application Download PDFInfo
- Publication number
- CN101614737A CN101614737A CN200910016309A CN200910016309A CN101614737A CN 101614737 A CN101614737 A CN 101614737A CN 200910016309 A CN200910016309 A CN 200910016309A CN 200910016309 A CN200910016309 A CN 200910016309A CN 101614737 A CN101614737 A CN 101614737A
- Authority
- CN
- China
- Prior art keywords
- virus
- coated
- antibody
- swine flu
- colloidal gold
- 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
Landscapes
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A kind of colloidal gold strip and its production and application, this test strips has an end liner, the sticking obedient coated film in this middle part above end liner, this coated film middle part have a bag by swine flu H1N1 virus detection of antibodies line and a bag by the control line of anti-mouse IgG antibody, the glass fibre membrane and the thieving paper of a sticking obedient coated with gold labeling antibody respectively about coated film top; Paste a sample pad above the glass fibre membrane of coated with gold labeling antibody.The present invention utilizes modern international state-of-the-art colloidal gold immunochromatographimethod technology, the colloidal gold strip of the detection swine flu H1N1 virus that a kind of terrain peasant household, basic unit usefulness such as test oneself is provided, this test strips has high specificity, highly sensitive, advantage such as detection speed is fast, and need not use instrument and equipment, with low cost, easy and simple to handle, can be widely used in terrain peasant household, basic unit and individual detection for the swine flu H1N1 virus.
Description
Technical field
The invention belongs to Preventive Veterinary Medicine check field, specifically relate to a kind of colloidal gold strip and its production and application.
Background technology
Swine influenza virus (swine influenza virus, SIV) belong to the A of orthomyxoviridae family type Influenza Virus, the pig that can cause different days, sex and kind is taken place acute, hot and height contact respiratory infectious disease, and it is a feature with burst, high heat, cough, expiratory dyspnea, depletion and death clinically.U.S.'s reported first in 1918 breaking out of swine flu, nineteen thirty shope has separated H1N1 hypotype swine influenza virus first from the pig body.Up to now, swine flu has spread all over U.S., Europe, Asia and Africa etc. all over the world.At present, the SIV that has been found that comprises hypotypes such as H1N1, H1N2, H1N7, H3N2, H3N6, H4N6, H5N1 and H9N2, but based on classical H1N1, bird H1N1 and class people H3N2 hypotype strain.China's swine flu in 1969 by the Taiwan reported first, from swinery, be separated to H1N1 hypotype swine influenza virus in 1991 first.Guo Yuanji (1985), Ni Hanzhong (2000), Zhang Suhua (2002), Li Haiyan (2003) etc. are to having done serosurvey in China different regions, and the result shows that with increasing progressively of time, China's swine flu positive rate increases.
At present, detection method to influenza virus, mainly containing virus separates, serodiagnosis and molecular biology method, but viral disengaging time is long, the requirement condition height, separation rate is low, and gather the patients acuity phase and convalescence paired sera carry out TPPA and confirm, can not in time diagnose again, therefore, these two kinds of methods in use are very restricted, and immune-gold labeled method (ImmunochromatographyAssay) is to grow up the early 1990s, quick because of it, easy and simple to handle, stable reagent, but room temperature accumulating, being difficult for pollution characteristics is widely used.It is the combination of immune affine technology, marking technology, immunolabelling technique and chromatographic technique.With coated antibody, colloid gold label antibody immobilization, combine with sample sorbing material etc., prepare the immunochromatography diagnostic test, only need insert sample solution to this strip down during use, several minutes just can judged result.With the immunity percolation method relatively, good stability operate easylier, quick, and owing to be dried strip form, need not cryopreservation, accumulating also makes things convenient for, and does not still at present see that about the detection of swine flu H1N1 virus available device or other testing tool are arranged.
Summary of the invention
The purpose of this invention is to provide a kind of colloidal gold strip and its production and application, can satisfy the demand of prior art.
A kind of colloidal gold strip, it is characterized in that an end liner is arranged, the sticking obedient coated film in this middle part above end liner, this coated film middle part has a bag to be done the control line of mouse IgG antibody by swine flu H1N1 virus detection of antibodies line and bag, the glass fibre membrane and the thieving paper of a sticking obedient coated with gold labeling antibody respectively about coated film top; Paste a sample pad above the glass fibre membrane of coated with gold labeling antibody.
The preparation method of above-mentioned colloidal gold strip, it is characterized in that preparation coated film earlier, to be cushioned the monoclonal antibody of swine flu H1N1 virus of liquid dilution or the polyclonal antibody and the anti-mouse IgG antibody dilution of anti-swine flu H1N1 virus with bag, and the two is sprayed on respectively on the nitrocellulose membrane middle part, form detection line and control line respectively, preparation is coated with the glass fibre membrane of deposit labeling antibody again, then coated film is sticked on the end liner, on coated film about the glass fibre membrane and the thieving paper of a sticking respectively obedient coated with gold labeling antibody; Paste a sample pad above the glass fibre membrane of coated with gold labeling antibody, make big plate, cutting promptly gets colloidal gold strip.
The application of above-mentioned colloidal gold strip in detecting the swine flu H1N1 virus.
The present invention utilizes modern international state-of-the-art colloidal gold immunochromatographimethod technology, the colloidal gold strip of the detection swine flu H1N1 virus that a kind of terrain peasant household, basic unit usefulness such as test oneself is provided, this test strips has high specificity, highly sensitive, advantage such as detection speed is fast, and need not use instrument and equipment, with low cost, easy and simple to handle, can be widely used in terrain peasant household, basic unit and individual detection for the swine flu H1N1 virus.
Description of drawings
Accompanying drawing is the vertical profile structural representation of test strips of the present invention.
Embodiment:
The preparation of colloidal gold strip
The preparation of the cell strain of monoclonal antibody of swine flu H1N1 virus and calibrating:
Slowly stir the chick embryo allantoic liquid that contains H1N1, and adding sodium chloride, making its final concentration is 0.5mol/L, adds the Macrogol 6000 (PEG6000) of 10% (percent by weight) again, 4 ℃ are spent the night, centrifugal 30 minutes of 8000r/min collects the virus precipitation, with phosphate buffer (PBS) dissolving of virus precipitation, 4 ℃ are spent the night, centrifugal 1 hour of 10000r/min, the gained precipitation is concentrated virus, is stored in-20 ℃ of low temperature refrigerators standby.
Take out after the viruses that concentrate dissolve from-20 ℃ of low temperature refrigerators, give BALB/C mice multiple spot hypodermic injection (0.5ml/ only), 15 days at interval, immunity was 3 times altogether, merged preceding 3 days, attacked with the antigen amount of above-mentioned concentrated viral 0.25ml at mouse peritoneal.With DMEM cultivation SP2/0 cell that contains 10% percent by volume calf serum and the lymphocyte of the BALB/C mouse of immunity, respectively by 2 * 10
7With 2 * 10
8Ratio merges with polyethylene glycol 1500 (PEG1500), fusion hole supernatant is added in the antigen coated tygon of swine flu H1N1 virus 96 holes respectively pulls, and detects with the ELISA method, determines the cell positive hole.To detect 9 positive strains of gained and carry out subclone, obtain 4 strains at last with limiting dilution assay.With the monoclonal antibody cell line of subclone gained in the external cultivation of going down to posterity, and carry out repeatedly liquid nitrogen cryopreservation and recovery, producing property swine flu H1N1 virus monoclonal antibody positive rate 100%, and the ability of maintenance stably excreting antibody, its ELISA tires and reaches more than 1: 1000.Every mouse peritoneal injecting fluid paraffin 0.5ml under aseptic condition, every the mouse peritoneal injection in a week back enlarged culture is 1-3 * IO
6The hybridoma of/0.2ml.Injection cell line after 7-10 days, or mouse once gathers ascites before dying, preserves down for-20 ℃.Adopt ammonium sulfate precipitation method slightly to carry, and then,, only show single protein band through the calibrating of SDS-PAGE electrophoresis with HITraprProteinA post purifying.Adopt the experiment of NaSCN competitive ELISA, measure its ED
50Drop-out value, reflect the size of its antibody affinity indirectly.Select wherein affinity 3 strains preferably, carry out hypotype calibrating, carry out the detection of Ig subclass with the monoclonal antibody of SRID pair cell culture supernatant, the subclass of above-mentioned 3 strain monoclonal antibodies is respectively: IgG1 has a strain; IgG2a has a strain; IgG2b has a strain.Detect with the antigen binding site of addition ELISA method the 3 strain monoclonal antibodies of non-lgM.The result shows in this 3 strain, three kinds of different antigen binding sites are arranged, and has excellent specificity for the swine flu H1N1 virus more.
The preparation of coated film 4:
Bag is cushioned the preparation of liquid: with the carbonate buffer solution (PBS) of 0.05M, pH9.6, use the 0.22u membrane filtration, put 4 ℃ standby, the term of validity 7 days.
The preparation of sealing damping fluid: with the PBS of 0.01M, pH7.0, with 0.22u membrane filtration mistake, put 4 ℃ standby, the term of validity 7 days.
Preparation sealing working fluid: will contain the PBS of 2%BSA, 2% skimmed milk, 0.01M, pH7.0, with 0.22u membrane filtration mistake, put 4 ℃ standby, the term of validity 3 days.
Detection line 6 preparation: debugging Membrane jetter, spouting liquid are 25 microlitres/35 centimetre, are cushioned the monoclonal antibody that liquid dilutes anti-pig sense H1N1 virus with bag, and concentration is 70ug/ml, rules on the nitrocellulose membrane middle part, and room temperature was dried 20 minutes.
Control line 7 preparation: debugging Membrane jetter, spouting liquid are 25 microlitres/35 centimetre, are cushioned liquid and dilute anti-mouse IgG antibody with wrapping, concentration is 2mg/ml, rules on nitrocellulose membrane, and this line is parallel with detection line, line-to-line should be careful even every 5mm, and room temperature was dried 20 minutes.The liquid that this two line is used infiltrates respectively in the nitrocellulose membrane, promptly is respectively detection line 6 and control line 7.
Coated film 4 preparations: the nitrocellulose membrane that will contain detection line 6 and control line 7 with above-mentioned sealing working fluid sealed 60 minutes for 37 ℃, took out rearmounted 37 ℃ of oven dry and handled two hours, and envelope is standby.
The manufacturing of the glass fibre membrane 3 of coated with gold labeling antibody
The preparation of gold chloride: the 10g gold chloride is made into 1% solution with 1000ml distilled water dissolving, put 4 ℃ standby, the term of validity 3 days.
The configuration of trisodium citrate: dissolve trisodium citrate with distilled water, be made into 1% solution, put 4 ℃ standby, the term of validity 3 days.
0.1M the preparation of sal tartari: 13.8g sal tartari is mixed with the 0.1M solution of potassium carbonate with the 1000ml distilled water, 0.22u membrane filtration mistake, put 4 ℃ standby, the term of validity 7 days.
The preparation of mark washing lotion: 20g BSA is configured to 2%BSA solution with 1000ml 0.01M, pH7.0 PBS, 0.22u membrane filtration mistake, put 4 ℃ standby, the term of validity 15 days.
The configuration that the gold labeling antibody is preserved liquid: with 10g BSA, 5g skimmed milk power, 0.5g NaN
3Dissolve in 1000ml 0.01M, pH 7.0 PBS with 1ml Tween-20, with 0.22u membrane filtration mistake, put 4 ℃ standby, the term of validity 15 days.
Firing of collaurum: 1% gold chloride is diluted to 0.01% with distilled water, put electric furnace and boil, add 4 milliliter of 1% citrate three sodium, continue to boil by per 100 milliliter of 0.01% gold chloride, be shiny red up to liquid and promptly stop heating, supply dehydration after being cooled to room temperature.Outward appearance should be pure, and is bright, do not have precipitation and floating thing.
The preparation of gold labeling antibody: transfer collaurum pH value to 7.6 with 0.1M sal tartari, the monoclonal antibody that adds the anti-swine flu H1N1 virus by 10 micrograms antibody/milliliter collaurum, mixing left standstill 30 minutes, centrifugal 30 minutes of 12000rpm, abandon supernatant, precipitation mark cleansing solution washed twice, last supernatant discarded will precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby, the term of validity 7 days.
The collaurum that mark is good is layered on the glass fibre membrane equably, 10 square centimeters of every ml soln shops, drying, envelope, put 4 ℃ standby.
The making of colloidal gold strip
Described end liner 1, sample pad 2 and thieving paper 5 are the general materials in this area.Above-mentioned coated film 4, the glass fibre membrane 3 that covers golden labeling antibody, end liner 1, sample pad 2 and thieving paper 5 are pasted successively, obtained test paper plate, the test strips that at last this test paper plate is cut into different in width gets final product.
The application of above-mentioned colloidal gold strip in detecting the swine flu H1N1 virus
When detecting in the blood sample swine flu H1N1 virus with this colloidal gold strip, if contain the swine flu H1N1 virus in the detection blood sample, this fluid drips is added on the sample pad 2 of this test strips, this liquid is subsequently when test strips enters in the glass fibre membrane 3 of coated with gold labeling antibody on the test strips, the monoclonal antibody of the swine flu H1N1 virus on the swine flu H1N1 virus that contains in the blood sample and this test strips in the glass fibre membrane 3 of coated with gold labeling antibody forms corresponding compound, move ahead with coated film 4 on the monoclonal antibody of swine flu H1N1 virus or the polyclonal antibody of anti-swine flu H1N1 virus form the aubergine lines, promptly form the aubergine band at detection line 6 places, continue to move ahead with coated film 4 in anti-mouse IgG antibody form the aubergine lines, promptly form the aubergine band at control line 7 places.If the aubergine band does not appear in control line 7, illustrate that then this test strips lost efficacy.If do not contain the swine flu H1N1 virus in the detection blood sample, then detection line 6 places the aubergine band can not occur, and the aubergine band must still appear in control line 7 places; If the aubergine band does not appear in control line 7, illustrate that then this test strips lost efficacy.
This test strips also can be used for detecting the fluid sample that contains in pig oral cavity or the anal swab, behind the centrifugal in oral cavity or the anal swab, the fluid drips of gained is added on the sample pad 2 of this test strips, and the result of appearance is identical with said process.
Claims (4)
1, a kind of colloidal gold strip, the sticking obedient coated film (4) in the middle part of it is characterized in that an end liner (1) is arranged, this end liner (1) is top, this coated film (4) middle part have a bag by swine flu H1N1 virus detection of antibodies line (6) and a bag by the control line (7) of anti-mouse IgG antibody, glue the glass fibre membrane (3) and the thieving paper (5) of an obedient coated with gold labeling antibody about coated film (4) top respectively; Paste a sample pad (2) above the glass fibre membrane of coated with gold labeling antibody (3).
2, colloidal gold strip according to claim 1 is characterized in that the antibody of the swine flu H1N1 virus of bag quilt in the described detection line (6) is highly purified anti-monoclonal antibody and/or polyclonal antibody.
3, the preparation method of the described colloidal gold strip of claim 1, it is characterized in that preparation coated film (4) earlier: will be cushioned the monoclonal antibody of swine flu H1N1 virus of liquid dilution or the polyclonal antibody and the anti-mouse IgG antibody dilution of anti-swine flu H1N1 virus with bag, and the two is sprayed on respectively on the nitrocellulose membrane middle part abreast, form detection line (6) and control line (7) respectively after infiltrating nitrocellulose membrane; Preparation is coated with the glass fibre membrane (3) of deposit labeling antibody again; Then coated film (4) is sticked on the middle part above the end liner (1), on coated film (4) about the glass fibre membrane (3) and the thieving paper (5) of a sticking obedient coated with gold labeling antibody respectively; Paste a sample pad (2) above the glass fibre membrane of coated with gold labeling antibody (3), make big plate, cutting promptly gets colloidal gold strip.
4, the application of the described colloidal gold strip of claim 1 in detecting the swine flu H1N1 virus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910016309A CN101614737A (en) | 2009-06-02 | 2009-06-02 | A kind of colloidal gold strip and its production and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910016309A CN101614737A (en) | 2009-06-02 | 2009-06-02 | A kind of colloidal gold strip and its production and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101614737A true CN101614737A (en) | 2009-12-30 |
Family
ID=41494490
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910016309A Pending CN101614737A (en) | 2009-06-02 | 2009-06-02 | A kind of colloidal gold strip and its production and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101614737A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101819204A (en) * | 2010-05-28 | 2010-09-01 | 青岛康地恩药业有限公司 | Collaurum test paper as well as preparation method and application thereof |
| CN101852802A (en) * | 2010-05-05 | 2010-10-06 | 青岛康地恩药业有限公司 | Test paper for detecting newcastle disease and infectious bronchitis viruses in one step |
| CN102565382A (en) * | 2012-01-06 | 2012-07-11 | 苏州浩欧博生物医药有限公司 | Immunochromatography method for detecting allergen-specific IgE antibodies in blood samples |
| CN102816865A (en) * | 2011-10-31 | 2012-12-12 | 武汉大学 | H1N1, PRRSV and CSFV quadruple detection kit |
| CN103336121A (en) * | 2013-06-13 | 2013-10-02 | 华中农业大学 | Swine influenza virus H3 subtype colloidal gold detection kit as well as preparation method and application thereof |
| CN110261605A (en) * | 2019-06-25 | 2019-09-20 | 陕西科技大学 | A kind of method of chitin modified enhancing test paper detection sensitivity |
| CN113189338A (en) * | 2021-04-30 | 2021-07-30 | 三峡大学 | Rhodamine B colloidal gold detection test strip and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101363865A (en) * | 2008-05-26 | 2009-02-11 | 北京庄笛浩禾生物医学科技有限公司 | Test paper strip for detecting PRRSV antibody colloidal gold, method for making same and applications |
| CN201397332Y (en) * | 2009-05-08 | 2010-02-03 | 天津市畜牧兽医研究所 | Colloidal gold test strip for rapidly detecting swine influenza virus |
-
2009
- 2009-06-02 CN CN200910016309A patent/CN101614737A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101363865A (en) * | 2008-05-26 | 2009-02-11 | 北京庄笛浩禾生物医学科技有限公司 | Test paper strip for detecting PRRSV antibody colloidal gold, method for making same and applications |
| CN201397332Y (en) * | 2009-05-08 | 2010-02-03 | 天津市畜牧兽医研究所 | Colloidal gold test strip for rapidly detecting swine influenza virus |
Non-Patent Citations (1)
| Title |
|---|
| 张祥斌等人: "H1N1亚型猪流感病毒HA基因的表达及单克隆抗体的制备", 《中国兽医科学》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101852802A (en) * | 2010-05-05 | 2010-10-06 | 青岛康地恩药业有限公司 | Test paper for detecting newcastle disease and infectious bronchitis viruses in one step |
| CN101819204A (en) * | 2010-05-28 | 2010-09-01 | 青岛康地恩药业有限公司 | Collaurum test paper as well as preparation method and application thereof |
| CN102816865A (en) * | 2011-10-31 | 2012-12-12 | 武汉大学 | H1N1, PRRSV and CSFV quadruple detection kit |
| CN102816865B (en) * | 2011-10-31 | 2014-04-02 | 武汉大学 | H1N1, PRRSV and CSFV quadruple detection kit |
| CN102565382A (en) * | 2012-01-06 | 2012-07-11 | 苏州浩欧博生物医药有限公司 | Immunochromatography method for detecting allergen-specific IgE antibodies in blood samples |
| CN102565382B (en) * | 2012-01-06 | 2013-12-04 | 苏州浩欧博生物医药有限公司 | Immunochromatography method for detecting allergen-specific IgE antibodies in blood samples |
| CN103336121A (en) * | 2013-06-13 | 2013-10-02 | 华中农业大学 | Swine influenza virus H3 subtype colloidal gold detection kit as well as preparation method and application thereof |
| CN103336121B (en) * | 2013-06-13 | 2015-03-04 | 华中农业大学 | Swine influenza virus H3 subtype colloidal gold detection kit as well as preparation method thereof |
| CN110261605A (en) * | 2019-06-25 | 2019-09-20 | 陕西科技大学 | A kind of method of chitin modified enhancing test paper detection sensitivity |
| CN113189338A (en) * | 2021-04-30 | 2021-07-30 | 三峡大学 | Rhodamine B colloidal gold detection test strip and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101614737A (en) | A kind of colloidal gold strip and its production and application | |
| CN102747040B (en) | Anti-influenza A virus nucleoprotein monoclonal antibody, its preparation and application | |
| CN104011077B (en) | From the binding molecule with influenza A virus neutralization activity that human B cell is produced | |
| CN102127523B (en) | Hybridoma cell line and application thereof | |
| CN107942061A (en) | A kind of test card, preparation and its detection method for detecting transmissible gastro-enteritis virus antibody | |
| JP5463107B2 (en) | Monoclonal antibodies that specifically differentiate new influenza and immunodetection reagents using them | |
| CN103173420B (en) | Hybridoma cell capable of secreting anti-cardiac troponin I monoclonal antibodies and applications thereof | |
| CN105745542B (en) | The assay method of influenza A virus | |
| CN103048459B (en) | Immune detection reagent for detecting respiratory syncytial virus | |
| CN102305859B (en) | Activated toxic antibody kit for detecting porcine reproductive and respiratory syndrome virus | |
| CN105319359B (en) | Streptococcus pneumoniae quantum dot immunochromatographic assay detection card and preparing method and application thereof | |
| CN102830229A (en) | Detection reagent of treponema pallidum IgM (Immunoglobulin M) antigen colloidal gold method and preparation method thereof | |
| CN109187967A (en) | A kind of detection simultaneously distinguishes O-shaped, the duplex rapid detection card of A type foot and mouth disease virus and preparation method thereof | |
| CN105601744A (en) | Recombinant antibody resisting influenza a virus nucleoproteins as well as preparation method and application of recombinant antibody | |
| CN104569408A (en) | Fenoterol colloidal gold detection card and preparation method thereof | |
| CN105753981B (en) | The immune chromatography reagent kit of anti-human Respiratory Syncytial Virus(RSV) N protein antibody and the application antibody | |
| JP2010261912A (en) | Immunology-detection method of human influenza virus h3 subtype | |
| CN101726597A (en) | Test paper for detecting avian influenza virus and preparation method thereof | |
| CN101082626A (en) | ELISA reagent kit for testing Asial type foot-and-mouth disease virus | |
| CN101852802A (en) | Test paper for detecting newcastle disease and infectious bronchitis viruses in one step | |
| CN103513035A (en) | Test strip and method for detecting aflatoxin M1 | |
| CN201413328Y (en) | Colloidal gold test paper strip for rapidly testing swine flu H1N1 virus | |
| CN201859147U (en) | Test strip for detecting Newcastle disease and infectious bursal disease virus in one step | |
| CN102305856A (en) | Mycoplasma pneumoniae IgM antibody colloid gold method detecting kit and preparation method thereof | |
| CN201397335Y (en) | A test paper strip for Colloidal gold test for detecting H5N1 subtype avian influenza virus rapidly |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent of invention or patent application | ||
| CB02 | Change of applicant information |
Address after: 266061 Shandong Province, Qingdao high tech park by the road No. 29 Shandong High Building Room 606 Applicant after: Qingdao KDN Pharmaceutical Co., Ltd. Co-applicant after: Qingdao Boite Biopharmaceutical Co., Ltd. Address before: 266061 Shandong Province, Qingdao high tech park by the road No. 29 Shandong High Building Room 606 Applicant before: Qingdao Continent Biotech Co., Ltd. Co-applicant before: Qingdao Boite Biopharmaceutical Co., Ltd. |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: APPLICANT; FROM: QINGDAO CONTINENT BIOTECH CO., LTD. TO: QINGDAO KDN PHARMACEUTICAL CO., LTD. |
|
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20091230 |