CN106701688A - Hybridoma cell strain 7G11 and antibody thereof - Google Patents
Hybridoma cell strain 7G11 and antibody thereof Download PDFInfo
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- CN106701688A CN106701688A CN201710029162.1A CN201710029162A CN106701688A CN 106701688 A CN106701688 A CN 106701688A CN 201710029162 A CN201710029162 A CN 201710029162A CN 106701688 A CN106701688 A CN 106701688A
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- 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
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
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- 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
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Abstract
The invention relates to a hybridoma cell strain 7G11 and an antibody thereof. The hybridoma cell strain 7G11 is delivered to the China Center for Type Culture Collection for collection on December 27, 2016, with the collection number of CCTCC NO: C201703. The hybridoma cell strain 7G11 can generate a monoclonal antibody capable of resisting streptospora acid, can specifically detect the streptospora acid and has important significance for detection of the streptospora acid.
Description
Technical field
The invention belongs to biological technical field, and in particular to hybridoma cell strain 7G11, further relate to by the hybridoma
The antibody that strain 7G11 is produced.
Background technology
Thin Alternaria alternata bacterium ketone acid (Tenuazonic acid, TA) be that rod method is mould, rice blast is mould, sorghum point is mould etc.
One of toxic metabolic products produced under specified conditions, are after alternariol (Alternariol, AOH), alternariol monomethyl ether
The mould toxin of a kind of rod method that (Alternariol monomethyl ether, AME) has found afterwards.TA has acute toxicity, Asia
Acute toxicity, potential carcinogenicity, cytotoxicity, cause dizzy mammal, salivation, vomiting heartbeat then occur and overrun, eat
Road and stomach large-area hemorrhage and circulatory failure, death.Because TA has a various biological characteristic, and it is the mould toxin of rod method
The maximum one kind of toxicity, studies have reported that TA can cooperate with enhancing other mycotoxins, with huge potential hazard, in recent years
Each side is increasingly caused to pay close attention to and study.
TA synthesizing artificial antigens and polyclonal antibody prepare existing part and report that such as river great waves utilize carbodlimide method
TA structures are modified, artificial antigen is prepared with bovine serum albumin coupling；The good grade of horse to TA derive drawing using oximate method
Enter to be coupled arm, be coupled with bovine serum albumin and hemocyanin, the immune BALB/c mouse of conjugate prepares polyclonal antibody；Yang Xingxing
TA is derived successively Deng using hydrazine hydrate and glyoxalic acid, synthesis nitrogen-containing hetero conjugated double bond coupling arm enters with bovine serum albumin
Row coupling obtains comlete antigen, and the polyclonal antibody of specific recognition TA derivatives is prepared by the way that new zealand white rabbit is immunized, and
Set up ELISA detection method.
Polyclonal antibody preparation time is short, preparation cost is low, but poorer than monoclonal antibody in terms of specificity, different batches
There is difference and the immune response of different animals between.Monoclonal antibody specificity is high, once target cell strain is screened, it is theoretical
On can just produce completely the same antibody, and the consistent antibody of substantial amounts of performance can be obtained by continuous culture method, produce
The differences between batches of product are small.The current preparation on TA monoclonal antibodies and products thereof exploitation is currently in blank stage, there is no and appoints
What is reported.
The content of the invention
In view of this, an object of the present invention is to provide a kind of hybridoma cell strain 7G11；The purpose of the present invention it
Two is the preparation method for providing hybridoma cell strain 7G11；The third object of the present invention is to provide by hybridoma cell strain
The monoclonal antibody that 7G11 is produced；The fourth object of the present invention is the preparation method for providing said monoclonal antibody；The present invention
The fifth purpose be provide monoclonal antibody application.
To reach above-mentioned purpose, the present invention provides following technical scheme：
The biological deposits numbering of the 1st, hybridoma cell strain 7G11, the hybridoma cell strain 7G11 is CCTCC NO：
C201703。
2nd, the preparation method of the hybridoma cell strain 7G11, TAO couplings hemocyanin immunogene is mixed with Freund's adjuvant
Immune mouse is closed, potency is then taken more than 1：The splenocyte of 10000 immune mouse is merged with myeloma cell SP2/0,
ELISA detections are carried out after being cultivated 10 days after fusion, hybridoma is obtained through screening subclone.
Preferably, splenocyte described in fusion and myeloma cell SP2/0 quantity ratio are 1：20.
3rd, the monoclonal antibody produced by hybridoma cell strain 7G11, the biological deposits of the hybridoma cell strain 7G11 are compiled
Number be CCTCC NO：C201703.
4th, the preparation method of the monoclonal antibody, ascites is prepared by hybridoma cell strain 7G11, is then carried out
ProteinA is purified, and obtains monoclonal antibody.
5th, application of the monoclonal antibody in the reagent for preparing detection TAO antigens.
Preferably, the monoclonal antibody is in the reagent for preparing the elisa competition laws thin Alternaria alternata bacterium ketone acid of detection
Using.
Preferably, the monoclonal antibody is in the reagent for preparing the elisa indirect methods thin Alternaria alternata bacterium ketone acid of detection
Using.
The beneficial effects of the present invention are：Hybridoma cell strain 7G11, the hybridoma cell strain is exempted from by TAO-KLH immunogenes
Epidemic disease mouse, is then merged the splenocyte of immune mouse with myeloma cell SP2/0, with TAO-BSA as antigen or chain
The thin Alternaria alternata bacterium ketone acid of the mould generation of lattice spore does the monoclonal antibody that competition antigen selection detection hybridoma is produced, screening
Obtain the hybridoma that antibody titer is high, specificity is good；The cell can secrete anti-TAO-KLH monoclonal antibodies, Neng Gouyong
In detection TAO antigens, significant is detected to thin Alternaria alternata bacterium ketone acid.
Brief description of the drawings
In order that the purpose of the present invention, technical scheme and beneficial effect are clearer, the present invention provides drawings described below and carries out
Explanation：
Fig. 1 is purifying protein SDS-PAGE electrophoresis result figures.
Biomaterial preservation
Hybridoma cell strain 7G11 send China typical culture collection center preservation in the present invention, and deposit number is CCTCC
NO:C201703, address is located at Wuhan, China Wuhan University, and preservation date is on December 27th, 2016, and Classification And Nomenclature is hybridoma
Cell line 7G11.
Specific embodiment
The preferred embodiments of the present invention will be described in detail below.
The reagent and consumptive material used in the present invention are as follows：Protein Marker (bronze object is biological), Balb/c mouse (are purchased from
Yangzhou University), Acr, Bis, Tris (are purchased from Sigma companies), and SDS (is purchased from Amresco companies), and TEMED (is purchased from BIO-RAD
Company), 0.22 μm of sterile filters and bag filter (are purchased from Millipore companies), and DMEM in high glucose culture medium is (public purchased from Gibco
Department), cell counting count board (is purchased from Improved Neubauer), and atoleine (bronze object is biological), PEG (is purchased from sigama), and 96 is thin
Born of the same parents' culture plate (is purchased from Costar), and it is pure that other reagents are domestic analysis.
Embodiment 1, animal immune and exempt from rear mice serum bioactivity
5 6~8 week old female BAl BIcs/c mouse are chosen, by haptens TAO coupling hemocyanin (Keyhole Limpet
Hemocyanin, KLH) TAO-KLH immunogenes and Freund's adjuvant mixed immunity, the μ g of hypodermic injection 100,2-3 weeks booster immunization
Once (first time immune original mixes with Freund's complete adjuvant, adds exempt to mix with incomplete Freund's adjuvant thereafter).Wherein, half
Antigen TAO draws after being derived to thin Alternaria alternata bacterium ketone acid (Tenuaconic acid, TA) by the hydrochloride of carboxymethyl azanol half
Enter linking arm and obtain.
Four exempt from after take a blood sample detection, determine that antiserum detects former potency, inspection for TAO-BSA by indirect ELISA method
Survey condition：
Envelope antigen：TAO-BSA detections are former, the thin Alternaria alternata bacterium ketone acid (competition antigen) of the mould generation of rod method；Coating
Concentration：5 μ g/mL, 100 μ l/ holes；Coating buffer solution：Phosphate buffer (PBS, pH 7.4)；Secondary antibody：Mountain sheep anti mouse-HRP, 1/
5000, testing result is as shown in table 1.
Table 1, antiserum and TAO-BSA Elisa results
Initial dilution:1:500
Result shows that 5 mouse ELISA potency are all higher than 1：10000, can continue to arrange mouse serum and the mould product of rod method
The competitive ELISA detection of raw thin Alternaria alternata bacterium ketone acid, as a result as shown in table 2.
Table 2, antiserum and the thin Alternaria alternata bacterium ketone acid of rod method mould generation competition Elisa results
Note：The thin Alternaria alternata bacterium ketone acid initial dilution of the mould generation of rod method：100ug startings, 3 times of dilutions
Result shows that substantially, final choice 1# mouse carry out cell fusion experiment to 1#, 2#, 5# mouse competition performance.
Embodiment 2, cell fusion
1st, prepared by myeloma cell
Fusion the last week, with the culture medium Amplification Culture SP2/0 cells of DMEM containing 10%FBS.When fusion, cell is covered with
About 6 bottles of T25 Tissue Culture Flasks, collect SP2/0 cells in 50mL centrifuge tubes, 1000rpm centrifugations 5min on the day of fusion.Abandon
Supernatant, then adds 20mL DMEM basal mediums, dispels cell and then counts.
2nd, prepared by splenocyte
Four Post-immunisation serum ELISA potency are 1：More than 10000 mouse, exempts from for 3 days before fusion eventually, and intraperitoneal injection resists
100 μ g of original.The fusion same day mouse to be merged of cervical dislocation euthanasia.Spleen is taken with 75% alcohol-pickled 5min. is aseptic,
Spleen is put into the interior culture dish for there are 10mL DMEM bases to cultivate.Take screen cloth to be put into another plate, spleen is transferred to
On screen cloth, spleen is ground with syringe heart.Add DMEM to screen cloth, rinse screen cloth, splenocyte is more collected into flat
In ware.During cell moved into 10mL centrifuge tubes, splenocyte is washed twice with the DMEM without serum, 1000rpm centrifugation 5min are collected
Splenocyte is counted.
3rd, cell fusion
Mixing myeloma cell and splenocyte, make myeloma cell with splenocyte quantity ratio 1：20 are advisable.Cell is put
To in 50mL centrifuge tubes, diluted with DMEM basal mediums, then 1000rpm centrifugations 5min, abandon supernatant, shake centrifuge tube makes carefully
Born of the same parents are uniform.It is slowly added to 0.8mL 50%PEG, reacts 90 seconds, be subsequently adding 20-30mL DMEM culture mediums and terminate PEG, melts
The cell of conjunction reacts 10 minutes in being put into 37 DEG C of water-baths, 1000rpm centrifugation 5min, abandons supernatant and is subsequently adding HAT DMEM cultures
Base.The cell of fusion is taped against in 96 orifice plates, per the μ L of hole 100, Tissue Culture Plate is then put into CO2Cultivated in incubator.Melt
Check within 4 days after conjunction, hybridoma cell clone rate has a small amount of cell fragment more than 50%, and cell growth state is good, fusion
ELISA selective mechanisms are proceeded by after 10 days.Testing conditions：Envelope antigen：TAO-BSA detections are former, the mould generation of rod method thin
Alternaria alternata bacterium ketone acid (competition antigen)；Coating concentration：5 μ g/mL, 100 μ l/ holes；Coating buffer solution：Phosphate buffer
(PBS,pH 7.4)；Secondary antibody：Mountain sheep anti mouse-HRP, 1/5000, as a result as shown in Table 3 and Table 4.
Cell conditioned medium and TAO-BSA Elisa results after table 3, fusion
1. number plate
2. number plate
3. number plate
4. number plate
5. number plate
6. number plate
Cell conditioned medium and the thin Alternaria alternata bacterium ketone acid Elisa results of the mould generation of rod method after table 4, fusion
| Cell line title | 1A2 | 1A5 | 1B5 | 1C4 | 1C8 | 1D4 | 1E1 | 1E2 | 1E4 |
| OD values | 3.102 | 2.0833 | 1.807 | 0.248 | 2.549 | 0.260 | 0.235 | 0.266 | 0.279 |
| Cell line title | 1E6 | 1F10 | 1F11 | 1G10 | 1G12 | 1H9 | 1H12 | 2A4 | 2A4 |
| OD values | 0.221 | 0.258 | 0.256 | 0.323 | 0.222 | 0.429 | 3.407 | 0.181 | 2.037 |
| Cell line title | 2B5 | 2C8 | 2D4 | 2D11 | 2F2 | 2F11 | 2G11 | 2H11 | 3A4 |
| OD values | 0.191 | 0.283 | 0.251 | 0.248 | 0.221 | 0.219 | 0.232 | 0.228 | 1.499 |
| Cell line title | 3A12 | 3B6 | 3F3 | 3F11 | 3G8 | 3H1 | 3H5 | 4A4 | 4A5 |
| OD values | 0.863 | 0.179 | 3.085 | 3.152 | 0.172 | 0.244 | 0.233 | 0.204 | 0.387 |
| Cell line title | 4A6 | 4B9 | 4B10 | 4D4 | 4E4 | 4F8 | 5C10 | 5E3 | 5E7 |
| OD values | 0.264 | 3.527 | 0.189 | 1.412 | 0.252 | 0.253 | 0.540 | 0.662 | 0.170 |
| Cell line title | 5G10 | 5H2 | 6C4 | 6C9 | 6E3 | 6H6 | Positive control | Competition hole | Negative control |
| OD values | 1.631 | 1.988 | 0.175 | 2.929 | 0.340 | 0.252 | 3.129 | 0.617 | 0.101 |
Embodiment 3, fusion screening and subclone
1st, fusion screening
In the previous day of detection, 5 μ g/mL antigens are coated with elisa plate with PBS, overnight, next day draws the μ of cell conditioned medium 100
L/ holes carry out ELISA detections according to ELISA results, judge that (sample well OD values/negative hole OD value >=2.1 item are judged in positive hole
Positive hole.The positive hole that inspection whole plate is detected is chosen with single track pipettor, second confirmation detection is carried out, the positive is further confirmed that
Hole, it is determined that after positive hole cell be subcloned.
2nd, it is subcloned
Cell in the positive hole of piping and druming, is counted, and N/4mL DMEM culture mediums are added in centrifuge tube, takes 100 μ l cell suspensions
To in centrifuge tube, blow it is even after stay 1mL, add DMEM to 4mL, blow even, stay 100 μ l (about 2 drop) in ttom of pipe.Add in centrifuge tube
DMEM to 5mL, drops to the first three rows of 96 orifice plates after mixing, 1.8~2ml or so is stayed per the dropper bottom of hole one, adds DMEM extremely
5mL, blow it is even after drop to the row of D, E, F tri- of 96 orifice plates, dripped per hole one, ttom of pipe stays 1.5~1.8m or so, adds DMEM to 2.8
~3mL or so, blow it is even after drop to G, H row of 96 orifice plates, dripped per hole one, examined under a microscope after 7-10 days, detecting has gram
The hole of grand growth, marks the hole of monoclonal, and the positive monoclonal cell of picking as far as possible is subcloned again, and detection is extremely
After 100% positive, choose monoclonal hole Amplification Culture singling.Choosing 10 plants of positive and competitive fusion cell lines carries out Ya Ke
Grand, fixed 10 plants of results are as follows after being screened through ELISA：
Envelope antigen：TAO-BSA detections are former, the thin Alternaria alternata bacterium ketone acid (competition antigen) of the mould generation of rod method；Coating
Concentration：5 μ g/mL, 100 μ L/ holes；Coating buffer solution：Phosphate buffer (PBS, pH 7.4)；Secondary antibody：Mountain sheep anti mouse-HRP, 1/
5000, as a result as shown in table 5.
Cell conditioned medium Elisa results after table 5, singling
| Cell line title | Indirect ELISA-OD is worth | Competitive ELISA-OD is worth |
| 3B3 | 2.924 | 0.712 |
| 3B4 | 2.950 | 0.752 |
| 3D12 | 2.950 | 1.079 |
| 4A1 | 2.899 | 0.895 |
| 4B9 | 2.876 | 0.883 |
| 4D5 | 3.236 | 0.846 |
| 4F4 | 3.345 | 0.231 |
| 4G1 | 3.412 | 1.383 |
| 7F11 | 3.412 | 0.524 |
| 7G11 | 3.236 | 0.450 |
According to the above results, selection 7G11 cell lines prepare ascites, because the cell line competition performance is preferably, growing way is most
Hurry up.Hybridoma cell strain 7G11 send China typical culture collection center preservation, and deposit number is CCTCC NO:C201703, ground
Location is located at Wuhan, China Wuhan University, and preservation date is on December 27th, 2016, and Classification And Nomenclature is hybridoma cell strain 7G11.
Embodiment 4, ProteinA antibody purifications and identification
By TAO-KLH 7G11 ascites, ProteinA purifying is carried out.Protein A sepharose medium is taken, chromatographic column is loaded, by abdomen
Water presses 1 with PBS:Slow loading after 1 mixing, after glycine elution buffer solution elution is used after antibody combination, that is, obtains required purifying
Antibody, carries out 4 DEG C of dialysed overnights in PBS immediately, the next day carry out purity, concentration and bioactivity.
Antibody indirect, competitive ELISA bioactivity, concrete operations are as follows：
(1) coating plate is designed according to experiment needs, and is marked on lath；
(2) it is coated with：TAO-BSA is detected that antigen is diluted by 5 μ g/mL with PBS coating buffers, after mixing in addition lath, often
The μ L of hole 100,4 DEG C of refrigerator overnights；
(3) envelope antigen：TAO-BSA detects antigen；
(4) it is coated with concentration：Diluted by 5 μ g/mL, 100 μ L/ holes；
(5) it is coated with buffer solution：Phosphate buffer (PBS, pH 7.4)；
(6) close：After coating is good, coating buffer is discarded, board-washing 3 times adds 200 μ l confining liquids (PBST+1%BSA) per hole,
37 DEG C of insulating box 1h, take out ELISA Plate, discard interior liquid, board-washing 1 time；
(7) primary antibody reaction：
Indirect ELISA：Antibody purification is by 1/500,2 times of dilutions, 37 DEG C of insulating box 1h；
Competitive ELISA：The thin μ g/mL of Alternaria alternata bacterium ketone acid 0.1 startings of the mould generation of rod method, 2 times of dilutions, per the μ of hole 50
L, synchronous to add antibody purification, antibody purification is by 1/1000 dilution, every hole 50 μ L, 37 DEG C of insulating box 1h；
(8) secondary antibody reaction
ELISA Plate is taken out, interior liquid, board-washing 3 times, to the ELIAS secondary antibody for adding 100 μ L to dilute in every hole, enzyme mark two is discarded
It is anti-：Mountain sheep anti mouse-HRP, 1/5000,37 DEG C of insulating box 1 hour.
(9) develop the color
ELISA Plate is taken out, interior liquid is discarded, board-washing 4 times first adds 100 μ L TMB nitrite ions, according to the depth of color per hole
Determine developing time, general 37 DEG C, 15min.
(10) terminating reaction
100 μ L 1M HCL solution are added per hole, terminating reaction is engraved in 450nm readings on ELIASA, OD values are more than
The corresponding dilution factor in hole of 2.1 times of the negative control OD value of setting, it is determined as the potency of the sample.
Result is as shown in table 6 and table 7.
The indirect Elisa results of table 6,7G11 antibody purifications
Note：Initial dilution:1:500；Potency is sample OD/ blank OD>=2.1 highest dilution
Drawn by testing result, the antibody titer of 7G11 is all in 512K or so.
Table 7,7G11 antibody purifications competition Elisa results
AC testing result shows that purified antibodies concentration is 0.4mg/mL；Correspondence antibody volume is 4.0mL, so
Its purity is detected using electrophoresis method afterwards, as a result as shown in Figure 1.Result shows that present invention antibody purity after purification is high, can use
In detection correspondence antigen.
Finally illustrate, preferred embodiment above is merely illustrative of the technical solution of the present invention and unrestricted, although logical
Cross above preferred embodiment to be described in detail the present invention, it is to be understood by those skilled in the art that can be
Various changes are made to it in form and in details, without departing from claims of the present invention limited range.
Claims (8)
1. hybridoma cell strain 7G11, it is characterised in that：The biological deposits numbering of the hybridoma cell strain 7G11 is CCTCC
NO：C201703.
2. the preparation method of hybridoma cell strain 7G11 described in claim 1, it is characterised in that：Haptens TAO coupling blood is blue
Protein immunogen and Freund's adjuvant mixed immunity mouse, then take potency more than 1：The splenocyte and bone of 10000 immune mouse
Myeloma cells SP2/0 is merged, and ELISA detections are carried out after being cultivated 10 days after fusion, and it is thin to obtain hybridoma through screening subclone
Born of the same parents；The haptens TAO thin Alternaria alternata bacterium ketone acid is derived by the hydrochloride of carboxymethyl azanol half after introduce linking arm and
.
3. preparation method according to claim 2, it is characterised in that：Splenocyte described in fusion and myeloma cell SP2/
0 quantity ratio is 1：20.
4. the monoclonal antibody for being produced by hybridoma cell strain 7G11, it is characterised in that：The life of the hybridoma cell strain 7G11
Thing deposit number is CCTCC NO：C201703.
5. the preparation method of monoclonal antibody described in claim 4, it is characterised in that：Hybridoma cell strain 7G11 is prepared into abdomen
Water, then carries out ProteinA purifying, obtains monoclonal antibody.
6. application of the monoclonal antibody described in claim 4 in the reagent for preparing the thin Alternaria alternata bacterium ketone acid of detection.
7. application according to claim 6, it is characterised in that：The monoclonal antibody is preparing the detection of elisa competition laws
Application in the reagent of thin Alternaria alternata bacterium ketone acid.
8. application according to claim 6, it is characterised in that：The monoclonal antibody is preparing the detection of elisa indirect methods
Application in the reagent of thin Alternaria alternata bacterium ketone acid.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107255716A (en) * | 2017-06-22 | 2017-10-17 | 华南农业大学 | The monoclonal antibody and its enzyme-linked immune detection method of a kind of tenuazonic acid |
| CN110804092A (en) * | 2019-11-25 | 2020-02-18 | 浙江中医药大学 | Monoclonal antibody for recognizing alternaria tenuis and hybridoma cell strain AtA8 thereof |
| CN110804094A (en) * | 2019-12-03 | 2020-02-18 | 浙江中医药大学 | Monoclonal antibody for identifying alternaria tenuis and hybridoma cell strain AtD2 thereof |
| CN110862452A (en) * | 2019-11-15 | 2020-03-06 | 浙江中医药大学 | Monoclonal antibody capable of recognizing alternaria and hybridoma cell strain AaA1 thereof |
| CN110862970A (en) * | 2019-12-05 | 2020-03-06 | 浙江中医药大学 | Monoclonal antibody for recognizing alternaria tenuis and hybridoma cell strain AtG7 thereof |
| CN110894232A (en) * | 2019-11-19 | 2020-03-20 | 浙江中医药大学 | Monoclonal antibody for recognizing alternaria tenuis and hybridoma cell strain AtA1 thereof |
| CN110894233A (en) * | 2019-11-19 | 2020-03-20 | 浙江中医药大学 | Monoclonal antibody capable of recognizing alternaria and hybridoma cell strain AaC2 thereof |
| CN110894234A (en) * | 2019-11-29 | 2020-03-20 | 浙江中医药大学 | Monoclonal antibody capable of recognizing alternaria and hybridoma cell strain AaC5 thereof |
| CN110922479A (en) * | 2019-12-10 | 2020-03-27 | 浙江中医药大学 | Monoclonal antibody for recognizing alternaria tenuis and hybridoma cell strain AtH9 thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102516148A (en) * | 2011-11-23 | 2012-06-27 | 华南农业大学 | Half antigen and antigen of tenuazonic acid and preparation method and application thereof |
| CN103487575A (en) * | 2013-09-02 | 2014-01-01 | 华南农业大学 | Chemiluminescent enzyme-linked immunosorbent assay kit of tenuazonic acid and application method thereof |
| CN105503689A (en) * | 2015-12-17 | 2016-04-20 | 西南大学 | Alternarin tenuazonic acid artificial antigen, polyclonal antibody, preparation method and application |
-
2017
- 2017-01-16 CN CN201710029162.1A patent/CN106701688B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102516148A (en) * | 2011-11-23 | 2012-06-27 | 华南农业大学 | Half antigen and antigen of tenuazonic acid and preparation method and application thereof |
| CN103487575A (en) * | 2013-09-02 | 2014-01-01 | 华南农业大学 | Chemiluminescent enzyme-linked immunosorbent assay kit of tenuazonic acid and application method thereof |
| CN105503689A (en) * | 2015-12-17 | 2016-04-20 | 西南大学 | Alternarin tenuazonic acid artificial antigen, polyclonal antibody, preparation method and application |
Non-Patent Citations (1)
| Title |
|---|
| 王雅丽等: "基于光谱法研究异细交链孢菌酮酸半抗原-抗体的相互作用", 《现代食品科技》 * |
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| CN110862452A (en) * | 2019-11-15 | 2020-03-06 | 浙江中医药大学 | Monoclonal antibody capable of recognizing alternaria and hybridoma cell strain AaA1 thereof |
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| CN110894234A (en) * | 2019-11-29 | 2020-03-20 | 浙江中医药大学 | Monoclonal antibody capable of recognizing alternaria and hybridoma cell strain AaC5 thereof |
| CN110804094A (en) * | 2019-12-03 | 2020-02-18 | 浙江中医药大学 | Monoclonal antibody for identifying alternaria tenuis and hybridoma cell strain AtD2 thereof |
| CN110804094B (en) * | 2019-12-03 | 2023-04-07 | 浙江中医药大学 | Monoclonal antibody for identifying alternaria tenuissima and hybridoma cell strain AtD2 thereof |
| CN110862970A (en) * | 2019-12-05 | 2020-03-06 | 浙江中医药大学 | Monoclonal antibody for recognizing alternaria tenuis and hybridoma cell strain AtG7 thereof |
| CN110862970B (en) * | 2019-12-05 | 2022-12-20 | 浙江中医药大学 | Monoclonal antibody for identifying alternaria tenuissima and hybridoma cell strain AtG7 thereof |
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