CN108948185A - The rabbit alliin antibody that alliin antigen and its immune response obtain - Google Patents
The rabbit alliin antibody that alliin antigen and its immune response obtain Download PDFInfo
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- CN108948185A CN108948185A CN201810785505.1A CN201810785505A CN108948185A CN 108948185 A CN108948185 A CN 108948185A CN 201810785505 A CN201810785505 A CN 201810785505A CN 108948185 A CN108948185 A CN 108948185A
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- alliin
- antigen
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- rabbit
- immune response
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- XUHLIQGRKRUKPH-GCXOYZPQSA-N Alliin Natural products N[C@H](C[S@@](=O)CC=C)C(O)=O XUHLIQGRKRUKPH-GCXOYZPQSA-N 0.000 title claims abstract description 89
- XUHLIQGRKRUKPH-UHFFFAOYSA-N S-allyl-L-cysteine sulfoxide Natural products OC(=O)C(N)CS(=O)CC=C XUHLIQGRKRUKPH-UHFFFAOYSA-N 0.000 title claims abstract description 89
- XUHLIQGRKRUKPH-DYEAUMGKSA-N alliin Chemical compound OC(=O)[C@@H](N)C[S@@](=O)CC=C XUHLIQGRKRUKPH-DYEAUMGKSA-N 0.000 title claims abstract description 89
- 235000015295 alliin Nutrition 0.000 title claims abstract description 89
- 239000000427 antigen Substances 0.000 title claims abstract description 52
- 102000036639 antigens Human genes 0.000 title claims abstract description 52
- 108091007433 antigens Proteins 0.000 title claims abstract description 52
- 241000283973 Oryctolagus cuniculus Species 0.000 title claims abstract description 26
- 230000028993 immune response Effects 0.000 title claims abstract description 14
- 239000002671 adjuvant Substances 0.000 claims abstract description 25
- 238000002347 injection Methods 0.000 claims abstract description 12
- 239000007924 injection Substances 0.000 claims abstract description 12
- 239000002994 raw material Substances 0.000 claims abstract description 3
- 210000002966 serum Anatomy 0.000 claims description 8
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 7
- 108010000912 Egg Proteins Proteins 0.000 claims description 7
- 102000002322 Egg Proteins Human genes 0.000 claims description 7
- 210000004369 blood Anatomy 0.000 claims description 7
- 239000008280 blood Substances 0.000 claims description 7
- 235000014103 egg white Nutrition 0.000 claims description 7
- 210000000969 egg white Anatomy 0.000 claims description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 6
- 240000002234 Allium sativum Species 0.000 claims description 5
- 235000004611 garlic Nutrition 0.000 claims description 5
- 108010058846 Ovalbumin Proteins 0.000 claims description 4
- 229940092253 ovalbumin Drugs 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 4
- 210000003462 vein Anatomy 0.000 claims description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 3
- 229910021529 ammonia Inorganic materials 0.000 claims description 3
- 230000000890 antigenic effect Effects 0.000 claims description 3
- 229940098773 bovine serum albumin Drugs 0.000 claims description 3
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 238000010254 subcutaneous injection Methods 0.000 claims description 2
- 239000007929 subcutaneous injection Substances 0.000 claims description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 claims 1
- 238000000605 extraction Methods 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 16
- 230000005847 immunogenicity Effects 0.000 abstract description 7
- 238000011160 research Methods 0.000 abstract description 6
- 230000003053 immunization Effects 0.000 abstract description 4
- 238000009826 distribution Methods 0.000 abstract description 3
- 238000001727 in vivo Methods 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 3
- 230000000144 pharmacologic effect Effects 0.000 abstract description 3
- 210000003743 erythrocyte Anatomy 0.000 description 14
- 210000004698 lymphocyte Anatomy 0.000 description 10
- 230000004520 agglutination Effects 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 238000002360 preparation method Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 4
- 239000002953 phosphate buffered saline Substances 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 239000012267 brine Substances 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- SACKVYGWWZDWQS-UHFFFAOYSA-N 5,5-diethyl-1,3-diazinane-2,4,6-trione;hydrochloride Chemical compound Cl.CCC1(CC)C(=O)NC(=O)NC1=O SACKVYGWWZDWQS-UHFFFAOYSA-N 0.000 description 2
- 239000005662 Paraffin oil Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- -1 amino, carboxyl Chemical group 0.000 description 2
- 229960000190 bacillus calmette–guérin vaccine Drugs 0.000 description 2
- 210000000601 blood cell Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 244000153158 Ammi visnaga Species 0.000 description 1
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 101710145634 Antigen 1 Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 102100031547 HLA class II histocompatibility antigen, DO alpha chain Human genes 0.000 description 1
- 101000866278 Homo sapiens HLA class II histocompatibility antigen, DO alpha chain Proteins 0.000 description 1
- 239000004166 Lanolin Substances 0.000 description 1
- 240000002114 Satureja hortensis Species 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229940039717 lanolin Drugs 0.000 description 1
- 235000019388 lanolin Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 230000008313 sensitization Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 125000000101 thioether group Chemical group 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 239000007762 w/o emulsion Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/77—Ovalbumin
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
This application discloses the rabbit alliin antibody that the alliin antigen immune response in biologic product technology field obtains, raw material includes alliin antigen and Freund's complete adjuvant, alliin antigen and Freund's complete adjuvant mixed in equal amounts are obtained into adjuvant antigen, it takes in alliin antigen injection to rabbit body, alliin antibody is obtained after immune response.The present invention solves the immunogenicity of alliin, and immunizing rabbit obtains alliin antibody, and alliin antibody can be used for alliin Concentration Testing, in vivo distribution and pharmacological mechanism research, all have important application value to alliin basic and clinic studies.
Description
Technical field
The invention belongs to the families that biologic product technology field more particularly to a kind of alliin antigen and its immune response obtain
Rabbit alliin antibody.
Background technique
Alliin is the water-soluble amino acids of garlic, and chemical property is stablized, the hair of experiment in vitro research in recent years
Existing, alliin has antimicrobial, antitumor, anti-oxidant, immunological regulation and anti-angiocardiopathy effect, and so far there are no, and clinic is ground
Study carefully report.
Low molecular compound does not have immunogenicity generally, after it is in conjunction with high-molecular compound, body can be activated to exempt from
Epidemic disease cell generates associated antibodies.Alliin is low molecule amino acids, itself does not have immunogenicity, but alliin
, can be in conjunction with protein molecule with amino, carboxyl and thioether group, whether conjugate has immunogenicity, and stimulation body produces
Raw antibody, has no relevant report.Research prepares alliin antibody, first has to that alliin is made to form comlete antigen, this is for being avoided
Alliin causes body generation alliin antibody to be equally of great significance during clinical use, and prepares alliin and resist
The basic necessary condition of body.Followed by animal selects, and rabbit is laboratory common experimental animal, unrestricted with injecting pathway,
The advantages such as animal origin, economic and practical, in subsequent research, experiment reagent is easy to get, such as anti-rabbit IgG antibody.
Alliin antibody can be used for alliin Concentration Testing, in vivo distribution and pharmacological mechanism research, to alliin
Basic and clinic studies all have important application value.
Summary of the invention
The rabbit alliin antibody that the invention is intended to provide a kind of alliin antigen and its immune response obtains, it is existing to solve
There is technology not aiming at the problem that the relevant immunizing antigen of alliin and antibody.
The technical scheme is that alliin antigen, including albumen and alliin, which made with albumen
The mixture being mixed to form for carrier and alliin.Above-mentioned albumen can may be vegetable protein for animal protein.
Preferably, the albumen includes bovine serum albumin(BSA) or ovalbumin.Bovine serum albumin(BSA) or ovalbumin are this
Common protein carrier is invented, main foundation is inexpensive and is easy to get.
Preferably, ovalbumin is egg white powder.Egg white powder is preferentially selected in the present invention.
The rabbit alliin antibody that alliin antigen immune response of the invention obtains, which is to pass through
The extract that will be obtained after the alliin antigenic stimulus rabbit immune response.
Specifically, raw material includes alliin antigen and Freund's complete adjuvant, by alliin antigen and Freund's complete adjuvant etc.
Amount is mixed to get adjuvant antigen, takes in alliin antigen injection to rabbit body, and alliin antibody is obtained after immune response.Freund assistant
Agent is the most common adjuvant so as to enhancing immunogenicity in current zoopery, it can be divided into Freund's complete adjuvant and Freund again
Freund's incomplete adjuvant.Incomplete Freund's adjuvant system finish (paraffin oil or vegetable oil) is mixed with emulsifier, when itself and antigen
Mixing becomes water-in-oil emulsion, is used for inoculation, mycobacteria (such as BCG vaccine of inactivation is added in Freund's incomplete adjuvant
Bacterium) then become Freund's complete adjuvant.
More specifically, take alliin antigen and adjuvant antigen spare, selective body focuses on the rabbit of 2~3kg, is spaced within every 7 days
Injection is primary, the adjuvant antigen of first time intracutaneous injection 2ml, the alliin antigen of second of subcutaneous injection 3ml, third time vein
Inject the alliin antigen of 1ml, the 4th intravenous injection 2ml alliin antigen, serum point after Culling heart blood after final injection 7 days
From rear acquisition alliin antibody.
When detecting to antibody, use " O " type human blood cell as carrier, in conjunction with forming sensitization after alliin
Red blood cell and sensitized lymphocyte, detection and identification alliin antibody.
The working principle of the invention and the utility model has the advantages that
1, the present invention solves the immunogenicity of alliin first, and immunizing rabbit obtains alliin antibody, is garlic ammonia
The basic and clinic studies of acid provide material base and technical guarantee.
2, alliin antibody can be used for alliin Concentration Testing, in vivo distribution and pharmacological mechanism research, to garlic ammonia
Sour basic and clinic studies all have important application value.
3, the present invention is preferential selects egg white powder to form milky complex precipitate in conjunction with alliin as carrier, makes
Alliin has immunogenicity, becomes comlete antigen.
4, immune preparation family's rabbit-anti alliin antibody is carried out to rabbit with this antigen binding Freund's complete adjuvant.
Detailed description of the invention
Fig. 1 is the display figure that sensitized erythrocyte immunofluorescent test detects alliin antibody；
Fig. 2 is the display figure that sensitized lymphocyte immunofluorescent test detects alliin antibody.
Specific embodiment
Below by the further details of explanation of specific embodiment:
The preparation of antigen
The preparation of alliin antigen: it is 100mg/ml alliin solution that the egg white of 10ml, which is added to 2ml mass concentration,
Stirring mixes well, and the visible milky alliin of naked eyes is presented and egg white conjugate precipitates, forms alliin antigen.
The preparation of adjuvant antigen: weighing lanolin 5g, takes paraffin oil 20ml, and high pressure sterilization is spare, is Freund incomplete adjuvant,
Freund incomplete adjuvant 10ml is taken, 60mg BCG vaccine is aseptically added dropwise, side edged is milled, and is Freund's complete adjuvant；
Then the alliin antigen of equivalent is added dropwise, same side edged is milled, and is formed stable suspension, is obtained adjuvant antigen.
Animal immune
Rabbit requires 2~3kg of weight, and inoculation is carried out by table 1.
Table 1 is rabbit immunization injecting program
| Frequency injection | It is spaced in date (day) | Approach | Antigenic property | Dosage of inoculation (ml) |
| For the first time | 7 | It is intradermal | Adjuvant antigen | 2 |
| Second | 7 | Subcutaneously | Alliin antigen | 3 |
| For the third time | 7 | Vein | Alliin antigen | 1 |
| 4th time | 7 | Vein | Alliin antigen | 2 |
Blood sampling: final injection is taken a blood sample after 7 days, carries out antibody test, and Culling heart blood after final injection 7 days injects blood
In sterile Kolle flask, it is put into 37 DEG C of environment, is transferred to after 30 minutes in 4 DEG C of refrigerator after waiting for blood to solidify, it is sterile after 24 hours
Under the conditions of separation serum it is spare, carry out alliin antibody test.
Alliin antibody test
Blood cell separation: " O " type people anticoagulation lymphocyte separation medium separates lymphocyte, 2000r/min centrifugation
20min, lymphocyte and red blood cell respectively with brine three times.
Sensitized erythrocyte preparation: PH9.6,0.1M barbital-hydrochloride buffer 20ml and 100mg/ml alliin solution
1ml is added packed red cells 1ml, is placed at room temperature for 1 hour, and 2000r/min is centrifuged 10min and precipitates red blood cell, brine
Three times, precipitating red blood cell normal saline is spare at 2% red cell suspension.It is cold simultaneously with 2% red cell suspension smear
Wind is dry, and cold acetone is fixed, and -20 DEG C of refrigerators save backup.
The preparation of sensitized lymphocyte antigen slide: above-mentioned isolated lymphocyte, precipitating retain 0.2ml with when removing supernatant
Liquid cell dispersion；1mlPH9.6 0.1M barbital-hydrochloride buffer and 100mg/ml alliin solution 0.2ml, room temperature is added
It places 1 hour, 2000r/min is centrifuged 10min and precipitates red blood cell, and brine three times, is precipitated with guarantor when removing supernatant
0.2ml liquid cell dispersion is stayed, cell sheet, cold wind drying are prepared, cold acetone is fixed, and -20 DEG C of refrigerators save backup.
Slide agglutination test: 10% sensitized erythrocyte one drop is added dropwise in the drop of immune serum one on glass slide, is added, uses toothpick
It mills mixing, shakes slide and observe erythrocyte agglutination situation, observation in every 2 minutes is primary, occurs agglutination in ten minutes for the positive.And
Replace immune serum with the rabbit anteserum and physiological saline that are immunized without alliin, is carried out as negative control and saline control same
Sample test.
Tube agglutination test: taking 10 test tubes, and row is on rack for test tube, and physiological saline 0.5ml is added in every pipe in addition to the first pipe,
Serum presses the first pipe original serum, 1:2,1:4,1:8 ..., and doubling dilution to the 9th pipe, the tenth pipe is saline control, and every pipe is added
2% sensitized erythrocyte 0.5ml observes result after placing 45 minutes under 37 DEG C of environment.
Immunofluorescent test: sensitized lymphocyte and sensitized erythrocyte antigen slide are taken out from refrigerator, is placed at room temperature for 5 points
Clock is added dropwise immune serum and is covered on sample, sets and put in 37 DEG C of environment in wet box 4 hours；It takes out and uses PH7.4,0.01M PBS punching
Wash clean is added dropwise goat-anti rabbit fluorescence antibody and covers sample, put in 37 DEG C of environment 4 hours in wet box, uses PH7.4,0.01M with taking out
PBS, that is, phosphate buffered saline (PBS) is rinsed well, cold wind drying, fluorescence microscopy microscopic observation result.
As a result:
Slide agglutination test is evident that erythrocyte agglutination, normal rabbits blood the results show that Rabbit Sera is visible
Cleer and peaceful physiological saline is showed no sensitized erythrocyte agglutination.
It is 1:64 that tube agglutination test, which detects antibody titer, and ratio is smaller, and available antibodies are more.
Under fluorescence microscope: the visible combination fluorescence antibody of antigen slide of sensitized erythrocyte preparation generates the red of autofluorescence
Cell, such as Fig. 1.Sensitized lymphocyte prepares antigen slide, and lymphocyte combination fluorescence antibody generates autofluorescence lymphocyte,
Such as Fig. 2, shines to represent and contain alliin antibody in above-mentioned cell.
It will be apparent to those skilled in the art that without departing from the structure of the invention, several deformations can also be made
And improvement, these also should be considered as protection scope of the present invention, these all will not influence the effect and patent that the present invention is implemented.
Claims (6)
1. alliin antigen, which is characterized in that including albumen and alliin, which is using albumen as carrier and garlic
The mixture that propylhomoserin is mixed to form.
2. alliin antigen according to claim 1, which is characterized in that the albumen includes bovine serum albumin(BSA) or egg white
Albumen.
3. alliin antigen according to claim 2, which is characterized in that the ovalbumin is egg white powder.
4. the rabbit alliin antibody that any alliin antigen immune response obtains according to claim 1~3, feature
It is, which is the extraction by will obtain after the alliin antigenic stimulus rabbit immune response
Object.
5. the rabbit alliin antibody that immune response according to claim 4 obtains, which is characterized in that raw material includes garlic ammonia
Alliin antigen and Freund's complete adjuvant mixed in equal amounts are obtained adjuvant antigen, take alliin by sour antigen and Freund's complete adjuvant
In antigen injection to rabbit body, alliin antibody is obtained after immune response.
6. the rabbit alliin antibody that immune response according to claim 5 obtains, which is characterized in that take alliin antigen
Spare with adjuvant antigen, selective body focuses on the rabbit of 2~3kg, and interval injection in every 7 days is primary, the assistant of first time intracutaneous injection 2ml
Agent antigen, the alliin antigen of second of subcutaneous injection 3ml, third time are injected intravenously the alliin antigen of 1ml, the 4th vein
2ml alliin antigen is injected, obtains alliin antibody after serum separation after Culling heart blood after final injection 7 days.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810785505.1A CN108948185B (en) | 2018-07-17 | 2018-07-17 | Alliin antigen and rabbit alliin antibody obtained by immune response of alliin antigen |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810785505.1A CN108948185B (en) | 2018-07-17 | 2018-07-17 | Alliin antigen and rabbit alliin antibody obtained by immune response of alliin antigen |
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Cited By (1)
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| CN109627332A (en) * | 2019-01-15 | 2019-04-16 | 深圳康体生命细胞技术有限公司 | A kind of immune antiboidy and preparation method thereof |
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