CN106397586B - specific yolk antibody for resisting dermatophytes and preparation method and application thereof - Google Patents
specific yolk antibody for resisting dermatophytes and preparation method and application thereof Download PDFInfo
- Publication number
- CN106397586B CN106397586B CN201610786928.6A CN201610786928A CN106397586B CN 106397586 B CN106397586 B CN 106397586B CN 201610786928 A CN201610786928 A CN 201610786928A CN 106397586 B CN106397586 B CN 106397586B
- Authority
- CN
- China
- Prior art keywords
- cell wall
- dermatophyte
- yolk antibody
- dermatophytes
- preparation
- 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.)
- Expired - Fee Related
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/14—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from fungi, algea or lichens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/12—Aerosols; Foams
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
- A61Q17/005—Antimicrobial preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/10—Washing or bathing preparations
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/02—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from eggs
-
- 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
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Dermatology (AREA)
- Birds (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the technical field of biology, and discloses a specific yolk antibody for resisting dermatophytes, and a preparation method and application thereof. The preparation method comprises the following steps: 1) selecting two main pathogenic bacteria of dermatophytosis, culturing the two main pathogenic bacteria of dermatophytosis, and respectively extracting cell wall proteins of bacteria; 2) preparing antigen immune hen with the protein obtained in the step 1), obtaining eggs in five times in total, and extracting and purifying the yolk antibody from the eggs; 3) the IgY is used to prepare the preparation for resisting dermatophyte. The yolk antibody for resisting dermatophytes, which is prepared by utilizing two cell wall proteins of dermatophyte pathogenic bacteria, has strong specificity, can effectively inhibit dermatophytes, and has no toxic or side effect; the method of the invention is easy for large-scale production, has low cost and simple process, and the obtained product is a very safe preparation for resisting dermatophytosis.
Description
Technical Field
the invention belongs to the field of biological pharmacy technology, and particularly relates to a specific yolk antibody for resisting dermatophytes, and a preparation method and application thereof.
Background
Fungi are widely present throughout the world, mainly in soil and on humans and animals. Among them, a common disease caused by fungal infection is skin tinea. For example: beriberi, psoriasis, etc. The dermatophytes penetrate into the dermis during the proliferation process to stimulate the nervous system of the dermis to cause the symptom of pruritus; the local tissue fluid is gathered, and the skin tissue is easy to be secondarily infected by bacteria after being damaged, so that inflammatory reaction is caused, and a series of skin tissue pathological phenomena caused by infecting dermatophytes are called dermatophyte diseases. Dermatophytosis is a common disease in humans, and at least 10-20% of the world population may be infected with these pathogens. In recent years, due to the wide application of antibiotics and immunosuppressants, the increasing incidence of immunodeficiency diseases and other factors, the onset of dermatophytosis tends to increase gradually, and the health care consciousness of people is improved, so that the dermatophytosis is increasingly emphasized.
The dermatophytosis includes tinea corporis, tinea manuum, tinea pedis, tinea cruris, tinea facialis, etc. Common dermatophytes are classified into three species, trichophyton, microsporum and epidermophyton. The trichophyton rubrum and the trichophyton mentagrophytes are two most common pathogenic strains, can infect the head, the trunk and the limbs of a human body, and are the most main skin tinea types infected by the human body. The two kinds of bacteria belong to trichophyton, and the fungi have similar structures, so the serum antibodies obtained by the two kinds of bacteria have cross property and weak specificity. In addition, passive immunization with specific antibodies is one of the effective methods for preventing and treating infectious diseases. Immunotherapy often requires large amounts of specific antibodies. The development of safe, high titer and inexpensive antibodies has been the focus of current research.
Klemperer in 1893 discovered that antibodies present in oocyte-producing chicken serum were transferred to the yolk and provided passive immune protection to chicks as major immunoglobulins during early embryonic development. Since this antibody is similar to a mammalian IgG molecule and is present in the yolk, it is called a yolk antibody (IgY). Since it is difficult to separate and purify IgY from yolk, only the immune system of chickens and the structural properties of IgY have been studied for a long time thereafter. Until 1980, Poison et al proposed PEG precipitation to make the separation of IgY from yolk simple and easy, and the application of IgY was extensively studied by scholars at home and abroad. IgY has a number of significant advantages: firstly, the egg laid by the immune hen is only collected and extracted without killing or sampling blood, and the egg is in accordance with the modern animal protection method; secondly, a small amount of antigen is used for immunizing poultry to obtain a large amount of specific IgY with uniform quality, the cost is low, and the yield is high; due to the fact that the distance of the germ line is greatly different, the IgY cannot generate cross serological reaction with the IgG; IgY has more stable properties and is convenient to store and apply; compared with IgG, the important performance of IgY is that it not only has the ability of combining with antigen, but also has obvious inhibition to the growth of antigen bacteria. In view of its unique immunochemical properties and suitability for the production of specific antibodies, IgY has a wide application prospect in the fields of immunotherapy and immunodiagnosis, and its passive immune function can be used for resisting virus and bacterial diseases, and has the potential of developing functional foods and new drugs.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention mainly aims to provide a preparation method of a specific yolk antibody for resisting dermatophytes.
Still another object of the present invention is to provide a specific yolk antibody against dermatophytes prepared by the above preparation method.
The present invention also provides the use of the specific yolk antibody against dermatophytes.
The purpose of the invention is realized by the following technical scheme:
A preparation method of a specific yolk antibody for resisting dermatophytes comprises the following steps:
(1) Selecting Trichophyton rubrum or Trichophyton mentagrophytes, performing solid culture for one week, and performing liquid culture;
(2) After liquid culture, washing and centrifuging, grinding the obtained hyphae by liquid nitrogen, carrying out ultrasonic crushing, and centrifuging to obtain cell walls; extracting cell wall protein of fungi by cold alkali extraction method, and centrifuging to obtain supernatant; dialyzing the supernatant to obtain dermatophyte cell wall protein antigen;
(3) Selecting 20-week-old laying hens; diluting the dermatophyte cell wall protein antigen into 1mg/mL dermatophyte cell wall protein antigen solution by using PBS buffer solution; mixing and emulsifying a dermatophyte cell wall protein antigen solution and Freund's complete adjuvant according to the volume ratio of 1:1, and injecting 0.5mL into each hen by adopting a subcutaneous multipoint injection immunization mode; two weeks after the first immunization, performing first boosting immunization, mixing and emulsifying a dermatophyte cell wall protein antigen solution with equivalent volume of Freund incomplete adjuvant, injecting 0.5mL of the antigen solution into each hen by adopting a subcutaneous multipoint injection immunization mode, performing boosting immunization once every 15 days for 4 times in total, collecting immune eggs from the first boosting immunization, and storing the eggs in a storage tank at 4 ℃;
(4) sterilizing the obtained immune eggs with alcohol, breaking the shell, collecting the yolk, mixing with frozen distilled water according to the volume ratio of 1:9, centrifuging to obtain water-soluble components, precipitating with PEG6000 to obtain yolk antibody dry powder, and storing at-20 deg.C.
The solid culture in the step (1) adopts an SDA solid culture medium; the liquid culture medium adopts an SDA liquid culture medium.
And (3) adding 10mM EDTA and 10mM PMSF to inhibit protease in the liquid nitrogen grinding process in the step (2).
And (3) extracting the IgY by a PEG6000 precipitation method, and purifying the IgY by a saturated ammonium sulfate solution, wherein the purification effect is good, and the obtained IgY is relatively pure.
a specific yolk antibody for resisting dermatophytes prepared by the above method is provided.
The application of the specific yolk antibody for resisting dermatophytes in preparing ointment, spray, hand sanitizer or wet paper towel for preventing and treating dermatophyte diseases has no toxic or side effect, and is safe and reliable. The skin tinea diseases include tinea manuum, tinea unguium, and tinea corporis.
Compared with the prior art, the invention has the following advantages and beneficial effects: the yolk antibody for resisting dermatophytes, which is prepared by utilizing two cell wall proteins of dermatophyte pathogenic bacteria, has strong specificity, can effectively inhibit dermatophytes, and has no toxic or side effect; the method of the invention is easy for large-scale production, has low cost and simple process, and the obtained product is a very safe preparation for resisting dermatophytosis.
Drawings
FIG. 1 shows the decrease and growth rule of IgY potency of cell wall protein of Trichophyton rubrum.
FIG. 2 shows the decrease and growth rule of IgY potency of trichophyton mentagrophytes cell wall protein.
FIG. 3 is an SDS-PAGE electrophoresis of IgY, wherein 1 is crude IgY of Trichophyton rubrum cell wall protein; 2 is IgY pure product of trichophyton rubrum cell wall protein; 3 is crude IgY of trichophyton mentagrophytes cell wall protein; 4 is IgY purified product of trichophyton mentagrophytes cell wall protein.
Detailed Description
the present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1: preparation of dermatophyte protein antigen
(1) Culturing Trichophyton rubrum and Trichophyton mentagrophytes in an SDA solid culture medium, inoculating into a liquid culture medium, culturing in a large scale, centrifuging, and washing with PBS to obtain a large amount of mycelia;
(2) Extraction of cell wall proteins: grinding the hyphae cultured in the step (1) by adopting liquid nitrogen, adding PBS for dissolution, adding 10mM EDTA and 10mM PMSF for inhibiting protease; breaking cell wall by using an ultrasonic cell crusher and a homogenizer, releasing hyphal cytoplasm, and performing centrifugal separation to obtain cell walls; adding 0.1N NaOH at 4 deg.C into the cell wall precipitate, stirring at 4 deg.C for 24 hr, centrifuging at 4 deg.C for 12000g for 10min to obtain filtrate, and neutralizing the filtrate with precooled 1mol/l HCl; dialyzing the filtrate with 50mM Tris-HCl for three days to obtain the dermatophyte cell wall protein antigen. Diluting the dermatophyte cell wall protein antigen into 1mg/mL dermatophyte cell wall protein antigen solution by using PBS buffer solution for later use.
Example 2: preparation of anti-dermatophyte egg yolk antibody
(1) Immunizing egg-laying hens with anti-dermatophytosis antigen
Selecting 12 laying hens of 20 weeks old; mixing and emulsifying the dermatophyte cell wall protein antigen solution obtained in the example 1 and Freund's complete adjuvant according to the volume ratio of 1:1, and injecting 0.5mL of the dermatophyte cell wall protein antigen solution into each hen in a subcutaneous multipoint injection immunization mode; two weeks after the first immunization, performing first boosting immunization, mixing and emulsifying a dermatophyte cell wall protein antigen solution with equivalent volume of Freund incomplete adjuvant, injecting 0.5mL to each hen by adopting a subcutaneous multipoint injection immunization mode, performing boosting immunization every 15 days, performing boosting immunization for 4 times in total, collecting immune eggs from the first boosting immunization, continuously collecting immune eggs for three months, numbering according to days, and storing at 4 ℃;
(2) Purification of specific yolk antibody against dermatophytosis
Sterilizing the obtained immune eggs with alcohol, breaking the shells of the eggs, removing membranes, collecting egg yolks, measuring the volume of the egg yolks, adding 3 times of 3 volume of the egg yolks of 3.5 percent of PEG6000 PBS solution, uniformly mixing, adjusting the pH value to 7.2, stirring for 40min, and storing at 4 ℃ overnight; centrifuging at 4 deg.C for 20min at 10000g, collecting supernatant, filtering with four layers of gauze to obtain filtrate, and measuring filtrate volume; adding PEG6000 powder into the filtrate to make the final concentration 12%, adjusting pH to 7.2, mixing well, stirring for 30 min; centrifuging at 4 deg.C for 20min at 10000g, and removing supernatant to obtain precipitate, i.e. IgY crude product.
Dissolving the precipitate with 4 deg.C 1mol/l PBS buffer (pH 7.2), centrifuging at 4 deg.C 10000g for 10min, collecting the upper layer, dripping saturated ammonium sulfate solution with the same volume as the filtrate, and standing at 4 deg.C overnight; centrifuging at 4 deg.C and 10000g for 10min to obtain precipitate, dissolving with PBS buffer (pH 7.2), adding 1/2 volume saturated ammonium sulfate solution, and standing at 4 deg.C for 2 hr; centrifuging at 4 deg.C and 10000g for 20min to obtain precipitate; vacuum freeze drying at-60 deg.C for 12 hr to obtain pure dry powder of yolk antibody with specificity for resisting dermatophytosis, and storing at-20 deg.C.
Example 3: potency assay for specific yolk antibody against dermatophytosis
And (3) detecting the titer of the yolk antibody obtained by immunizing various antigens by ELISA. Adding 100 mul of whole-fungus antigens coated with trichophyton rubrum and trichophyton mentagrophytes into each hole of a 96-hole enzyme label plate, covering the whole-fungus antigens on the two enzyme label plates by using a sealing film, incubating for 2h at 37 ℃, then spin-drying, washing for 3 times by using PBST, and patting dry by using absorbent paper; adding 100 μ l/well of 30mg/mL skimmed milk powder, sealing, incubating at 37 deg.C for 1h, spin-drying, washing with PBST for 3 times, and drying with absorbent paper; diluting IgY obtained by immunizing two cell wall proteins of trichophyton rubrum and trichophyton mentagrophytes respectively in multiple ratios, adding 100 mu l/hole into an enzyme label plate, adding nonimmune in the last line as a blank control, incubating at 37 ℃ for 1h, spin-drying, washing with PBST for 3 times, and patting dry with absorbent paper; adding 100 mul of rabbit anti-chicken IgY-HRP antibody with the volume being 1:5000 into each hole, incubating for 1h at 37 ℃, spin-drying, washing for 3 times by PBST, and patting dry by absorbent paper; the reaction was stopped by adding 100. mu.l of TMB substrate per well, incubating at 37 ℃ for 15min, and then adding 50. mu.l of 2mol/l H2SO4 per well. The OD value at 490nm was measured by a microplate reader. The results are shown in fig. 1, fig. 2 and table 1.
TABLE 1 results of potency assay of specific yolk antibody against dermatophytosis
example 4: purity of specific yolk antibody against dermatophytosis determined by SDS-PAGE
the purity of the yolk antibody against dermatophytosis was checked by SDS-PAGE electrophoresis, as shown in FIG. 3. The method comprises the following specific steps: 12% separation gel and 5% concentrated gel are used, a marker is used as a reference substance, and after electrophoresis is finished, Coomassie brilliant blue is used for staining, and finally, decoloration is carried out. The results show that: the purity can reach more than 90%.
Example 5: in vitro bacteriostasis test
(1) Preparation of bacterial suspension: washing Trichophyton rubrum and Trichophyton mentagrophytes with PBS buffer solution, diluting, mixing the bacterial suspensions, and selecting 3 × 104 bacterial suspension by blood plate counting method.
(2) Bacteriostatic experiments: dissolving a proper amount of prepared IgY freeze-dried powder into a sterilized SDA liquid culture medium to enable the initial concentration to be 80mg/mL, and diluting the solution with the culture medium to obtain: 40,20,10,5,2.5,1.25mg/mL, 2mL each in 7 small tubes, 2mLSDA liquid culture medium based on the eighth tube as blank control. Then 2mL of the bacterial suspension is added. The mixture was incubated at 28 ℃ for one week on a shaker at 160 r/min. After the test tubes are taken out, a large number of mycelial pellets can be observed to grow on the blank control, and other test tubes have different degrees of inhibition effects on fungi. The diluted test tube with the antibody concentration of 20mg/mL is the minimum inhibitory concentration.
the above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (3)
1. A preparation method of a specific yolk antibody for resisting dermatophytes is characterized by comprising the following steps:
(1) Selecting Trichophyton rubrum or Trichophyton mentagrophytes, performing solid culture for one week, and performing liquid culture; the solid culture adopts an SDA solid culture medium; the liquid culture medium adopts an SDA liquid culture medium;
(2) after liquid culture, washing and centrifuging, grinding the obtained hyphae by liquid nitrogen, carrying out ultrasonic crushing, and centrifuging to obtain cell walls; extracting cell wall protein of fungi by cold alkali extraction method, and centrifuging to obtain supernatant; dialyzing the supernatant to obtain dermatophyte cell wall protein antigen; adding 10mM EDTA and 10mM PMSF to inhibit protease in the liquid nitrogen grinding process;
(3) Selecting 20-week-old laying hens; diluting the dermatophyte cell wall protein antigen into 1mg/mL dermatophyte cell wall protein antigen solution by using PBS buffer solution; mixing and emulsifying a dermatophyte cell wall protein antigen solution and Freund's complete adjuvant according to the volume ratio of 1:1, and injecting 0.5mL into each hen by adopting a subcutaneous multipoint injection immunization mode; two weeks after the first immunization, performing first boosting immunization, mixing and emulsifying a dermatophyte cell wall protein antigen solution with equivalent volume of Freund incomplete adjuvant, injecting 0.5mL of the antigen solution into each hen by adopting a subcutaneous multipoint injection immunization mode, performing boosting immunization once every 15 days for 4 times in total, collecting immune eggs from the first boosting immunization, and storing the eggs in a storage tank at 4 ℃;
(4) Sterilizing the obtained immune eggs with alcohol, breaking the shell, collecting the yolk, mixing with frozen distilled water according to the volume ratio of 1:9, centrifuging to obtain water-soluble components, precipitating with PEG6000 to obtain yolk antibody dry powder, and storing at-20 deg.C.
2. a specific yolk antibody against dermatophytes prepared according to the method of claim 1.
3. The use of the specific yolk antibody against dermatophytes according to claim 2 in the preparation of an ointment, a spray, a hand sanitizer or a wet wipe for preventing and treating dermatophyte diseases.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610786928.6A CN106397586B (en) | 2016-08-31 | 2016-08-31 | specific yolk antibody for resisting dermatophytes and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610786928.6A CN106397586B (en) | 2016-08-31 | 2016-08-31 | specific yolk antibody for resisting dermatophytes and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106397586A CN106397586A (en) | 2017-02-15 |
| CN106397586B true CN106397586B (en) | 2019-12-06 |
Family
ID=58000489
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610786928.6A Expired - Fee Related CN106397586B (en) | 2016-08-31 | 2016-08-31 | specific yolk antibody for resisting dermatophytes and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106397586B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108531468B (en) * | 2018-02-24 | 2020-07-17 | 广东工业大学 | Gene for coding recombinant helicobacter pylori hypha ammonia protease, yolk antibody and application |
| CN109010823B (en) * | 2018-08-27 | 2022-03-18 | 广州汇高生物科技有限公司 | Composition for resisting various pathogenic bacteria, preparation process and application thereof, spray and preparation process thereof |
| CN109575137A (en) * | 2018-12-11 | 2019-04-05 | 江南大学 | A kind of alpha-amylase inhibitor and its preparation method and application |
| CN116284366A (en) * | 2023-05-25 | 2023-06-23 | 南京黎明生物制品有限公司 | Quick detection antibody and kit for dermatophytosis and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1569896A (en) * | 2003-07-18 | 2005-01-26 | 广州新崴生物医药科技有限公司 | Preparation of composite specific yelk antibody igy against cutaneous infection and its use in treatment of cutaneous infection |
| CN104892756A (en) * | 2015-06-18 | 2015-09-09 | 成都安蒂康生物科技有限公司 | Antibacterial chicken egg yolk complex antibody as well as preparation method and application thereof |
| CN105106953A (en) * | 2015-08-21 | 2015-12-02 | 天津耀宇生物技术有限公司 | Antibody-based drug for treating psoriasis |
| JP2016034975A (en) * | 2011-08-19 | 2016-03-17 | オーストリッチファーマ株式会社 | Antibody and antibody-containing composition |
-
2016
- 2016-08-31 CN CN201610786928.6A patent/CN106397586B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1569896A (en) * | 2003-07-18 | 2005-01-26 | 广州新崴生物医药科技有限公司 | Preparation of composite specific yelk antibody igy against cutaneous infection and its use in treatment of cutaneous infection |
| JP2016034975A (en) * | 2011-08-19 | 2016-03-17 | オーストリッチファーマ株式会社 | Antibody and antibody-containing composition |
| CN104892756A (en) * | 2015-06-18 | 2015-09-09 | 成都安蒂康生物科技有限公司 | Antibacterial chicken egg yolk complex antibody as well as preparation method and application thereof |
| CN105106953A (en) * | 2015-08-21 | 2015-12-02 | 天津耀宇生物技术有限公司 | Antibody-based drug for treating psoriasis |
Non-Patent Citations (3)
| Title |
|---|
| 抗白色念珠菌鸡卵黄抗体(IgY)的研制;常山;《第三军医大学学报》;20020930;第24卷(第9期);全文 * |
| 抗须癣毛癣菌细胞壁蛋白的卵黄抗体制备和鉴定;胡青青;《中国免疫学杂志》;20171231;第33卷;全文 * |
| 真菌菌丝细胞壁可溶性蛋白抽提方法研究;张成省;《山东科学》;20040331;第17卷(第1期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106397586A (en) | 2017-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106397586B (en) | specific yolk antibody for resisting dermatophytes and preparation method and application thereof | |
| CN107656066A (en) | A kind of duck tembusu virus E protein truncated protein and application | |
| CN102406931A (en) | Pandemic influenza virus split vaccine | |
| CN108721615B (en) | A kind of method and its vaccine preparing duck tembusu virus inactivated vaccine | |
| CN104877968B (en) | Efficient secretion anti-dog parvovirus monoclonal antibody hybridoma cell A135 strains | |
| Gao et al. | Construction of a camelid VHH yeast two-hybrid library and the selection of VHH against haemagglutinin-neuraminidase protein of the Newcastle disease virus | |
| CN103059132A (en) | Monoclonal antibody of anti-H9 subtype flu virus haemagglutinin protein and application thereof | |
| CN107435041A (en) | Chimeric the newcastle Disease poisonous carrier H9 live vaccines Candidate Strain and its construction method for overcoming chicken Newcastle disease maternal antibody to influence | |
| CN104312979A (en) | Anti-H7 subtype avian influenza virus monoclonal antibody and application thereof | |
| AU646522B2 (en) | Diagnosis and treatment of multiple sclerosis | |
| CN104606675A (en) | Preparation method of poultry multi-linked specific egg yolk antibodies and transfer factors | |
| CN105198989A (en) | Shewanella-smarisflavi-resistant egg yolk antibody and preparation method thereof | |
| CN107831306B (en) | H7 subtype avian influenza virus double-antibody sandwich ELISA kit and detection method thereof | |
| JP2013147471A (en) | Aqueous solution of ostrich antibody | |
| CN106279349B (en) | extraction method of dermatophyte cytoplasm protein antigen for preparing yolk antibody and preparation method of yolk antibody | |
| CN104845941B (en) | A kind of avian infectious bronchitis virus IBV K136, cell strain of monoclonal antibody 3D5, monoclonal antibody and its application using its preparation | |
| KR101996736B1 (en) | Cell line derived from porcine kidney for mass production of Sacbrood virus and uses thereof | |
| US4666851A (en) | Specific mycoplasma membrane antigen and antibody and their clinical applications | |
| CN112143714B (en) | Method for producing H7 subtype avian influenza virus inactivated vaccine by using low-immunity chick embryo | |
| CN104357406A (en) | Rabies virus SNK-CTN strain and application thereof | |
| CN107384875A (en) | Chimeric the newcastle Disease poisonous carrier H7 live vaccines Candidate Strain and its construction method for overcoming chicken Newcastle disease maternal antibody to influence | |
| KR20180023246A (en) | Monoclonal antibody against nervous necrosis virus and use thereof | |
| CN108330101B (en) | Hybridoma cell strain, monoclonal antibody produced by hybridoma cell strain and application of monoclonal antibody | |
| CN111777684A (en) | Preparation method and application of antibody induced by coronavirus tandem epitope protein | |
| CN110172096A (en) | A kind of preparation method of Riemerlla anatipestifer monoclonal antibody |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20191206 Termination date: 20210831 |