CN104694482A - Hybridoma cell strain McAb 1G2 and rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody - Google Patents
Hybridoma cell strain McAb 1G2 and rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody Download PDFInfo
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Abstract
The invention provides a hybridoma cell strain McAb 1G2 and a rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody, and relates to the hybridoma cell strain and the monoclonal antibody secreted by the hybridoma cell strain. The hybridoma cell strain McAb 1G2 and the rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody solve the problem that no monoclonal antibody capable of recognizing RHDV2 type virus or RHDVa type virus independently exists. The preservation number of the hybridoma cell strain McAb 1G2 is CGMCCNo.10316. The rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody is generated by the hybridoma cell strain McAb 1G2 with the preservation number being CGMCC No.10316. By means of the rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody, whether rabbits suffering the rabbit hemorrhagic disease are specifically infected with the RHDV2 type virus or not can be recognized precisely, more targeted measures can be taken, and the hybridoma cell strain McAb 1G2 and the rabbit hemorrhagic disease RHDV2 type virus monoclonal antibody have far-reaching significance in the later immunization and prevention and curing.
Description
Technical field
The present invention relates to the monoclonal antibody of a strain of hybridoma strain and secretion thereof.
Background technology
Rabbit haemorrhagic disease (Rabbit Hemorrhagic Disease, RHD), be commonly called as " rabbit pest " (Rabbit Plague), by rabbit hemorrhagic disease virus (Rabbit Hemorrhagic Disease Virus, RHDV) acute, strong, the contagious disease caused, be changed to principal character with hepatic necrosis, substantial viscera oedema, respiratory system and whole body multiple organ extravasated blood and hemorrhagic, destructive strike is caused usually to rabbit keeping.
The people such as Ghislaine Le Gall-Recul é in 2013 report the appearance of RHDV2 C-type virus C, and the homology of RHDV2 C-type virus C and RHDVa C-type virus C Nucleocapsid Protein Gene sequence is 82.4%, and the homology of aminoacid sequence is 89.2%.The immunological cross protection ratio of this two strain virus is 75%.VP60 gene is viral unique capsid protein encoding gene, and in RHDVa C-type virus C, the homology of VP60 gene is between 93.7% ~ 99.8%; In RHDV2 C-type virus C, the homology of VP60 gene is 98.5%.Because the homology of RHDV2 C-type virus C and RHDVa C-type virus C VP60 protein amino acid sequence is 89.2%, both similar antigen sites are many.So, also there is not the monoclonal antibody that can identify separately RHDV2 C-type virus C or RHDVa C-type virus C at present.
Summary of the invention
The present invention is to solve the problem not having at present to identify separately the monoclonal antibody of RHDV2 C-type virus C or RHDVa C-type virus C, and the hybridoma cell strain McAb 1G2 provided and rabbit haemorrhagic disease RHDV2 C-type virus C monoclonal antibody.
Hybridoma cell strain McAb 1G2 of the present invention is mouse hybridoma cell (Mus musculus Hybridoma) McAb1G2, and be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.10316.
The monoclonal antibody of rabbit haemorrhagic disease RHDV2 C-type virus C of the present invention, the hybridoma cell strain McAb 1G2 being CGMCC No.10316 by preserving number produces.
The monoclonal antibody of the rabbit haemorrhagic disease RHDV2 C-type virus C that hybridoma cell strain McAb 1G2 of the present invention secretes can identify separately RHDV2 C-type virus C and VP602 albumen; This monoclonal antibody molecule heavy chain is IgG2a hypotype, and light chain is Kappa hypotype (as shown in Figure 2).
Whether what the rabbit utilizing the monoclonal antibody of rabbit haemorrhagic disease RHDV2 C-type virus C of the present invention to identify accurately to suffer from rabbit haemorrhagic disease specifically infected is RHDV2 C-type virus C, can take measures more targetedly, to the immunity in later stage and control, there is profound significance.
Hybridoma cell strain McAb 1G2 is mouse hybridoma cell McAb 1G2, belong to Mus (Mus), be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, preserving number is CGMCC No.10316, and preservation date is on January 21st, 2015.
Accompanying drawing explanation
Fig. 1 is the monoclonal antibody molecule hypotype qualification result figure of the rabbit haemorrhagic disease RHDVa C-type virus C that hybridoma cell strain McAb 1H2 secretes.
Fig. 2 is the monoclonal antibody molecule hypotype qualification result figure of the rabbit haemorrhagic disease RHDV2 C-type virus C that hybridoma cell strain McAb 1G2 secretes.
Fig. 3 is the ELISA qualification result figure of the antibody molecule secreted by hybridoma cell strain McAb 1H2, and in Fig. 3, a is RHDVa virus particle; B is restructuring Bac-VP60a albumen; C is restructuring Bac-VP602 albumen.
Fig. 4 is the ELISA qualification result figure of the antibody molecule secreted by hybridoma cell strain McAb 1G2, and in Fig. 4, a is RHDVa virus particle; B is restructuring Bac-VP60a albumen; C is restructuring Bac-VP602 albumen.
Fig. 5 is the Western blot qualification result figure of the antibody molecule secreted by hybridoma cell strain McAb 1H2, and in Fig. 5, M is protein molecular weight marker; B is restructuring Bac-VP60a albumen; C is restructuring Bac-VP602 albumen.
Fig. 6 is the Western blot qualification result figure of the antibody molecule secreted by hybridoma cell strain McAb 1G2, and in Fig. 6, M is protein molecular weight marker; B is restructuring Bac-VP60a albumen; C is restructuring Bac-VP602 albumen.
Fig. 7 is the IFA qualification result figure of the antibody molecule secreted by hybridoma cell strain McAb 1H2, and in Fig. 7, b is the Sf9 cell that rBacmid-VP60a infects; C is the Sf9 cell that rBacmid-VP602 infects.
Fig. 8 is the IFA qualification result figure of the antibody molecule secreted by hybridoma cell strain McAb 1G2, and in Fig. 8, b is the Sf9 cell that rBacmid-VP60a infects; C is the Sf9 cell that rBacmid-VP602 infects.
Embodiment
Technical solution of the present invention is not limited to following cited embodiment, also comprises the arbitrary combination between each embodiment.
Embodiment one: present embodiment hybridoma cell strain McAb 1G2 is mouse hybridoma cell (Mus musculus Hybridoma) McAb 1G2, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.10316, and preservation date is on January 21st, 2015.
Embodiment two: the monoclonal antibody of present embodiment rabbit haemorrhagic disease RHDV2 C-type virus C, the hybridoma cell strain McAb 1G2 being CGMCC No.10316 by preserving number produces.
Embodiment 1
1.1 prepare detectable antigens
RHDVa-VP60 gene (GenBank accession number:JF412629) and RHDV2-VP60 gene (GenBank accession number:HE800532) are cloned in pFastBacHTa carrier respectively, obtain recombinant plasmid pfast-VP60a and pfast-VP602, transformation of E. coli DH10Bac competent cell.Be inoculated into LB agar plate (containing kantlex 50mg/mL, gentamicin 40mg/mL, tsiklomitsin 50mg/mL, X-gal 50mg/mL and IPTG 142.8mg/mL), cultivate about 48h for 37 DEG C.Picking white colony, again streak culture on LB agar plate (containing kantlex 50mg/mL, gentamicin 40mg/mL, tsiklomitsin 50mg/mL, X-gal 50mg/mL and IPTG 142.8mg/mL), bacterium colony is defined as white person further and is inoculated in LB substratum (this LB substratum is containing kantlex 50mg/mL, gentamicin 40mg/mL and tsiklomitsin 50mg/mL), and 24 ~ 36h is cultivated in 37 DEG C of shakings.Extraction Bacmid is template, is about the fragment of 4.2kb with universal primer M13 (+) and M13 (-) pcr amplification length, is about the fragment of 4.0kb with primer M13 (+) and VP60 downstream primer pcr amplification length.Selection amplified fragments meets the restructuring Bacmid expecting that size, specificity are good, called after rBacmid-VP60a and rBacmid-VP602.
Infect Sf9 (SpAoptera frugiperda) cell with rBacmid-VP60a and rBacmid-VP602, synthesize specific restructuring Bac-VP60a and Bac-VP602 albumen.And using the detectable antigens of Bac-VP60a and Bac-VP602 albumen as monoclonal antibody of recombinating.
1.2 monoclonal antibody preparation
1.2.1 immune mouse experiment
The aseptic PBS substratum of recombinant protein Bac-VP60a after purifying and recombinant protein Bac-VP602 is diluted to 500 μ g/mL, after adding the Fu Shi Freund's complete adjuvant emulsification of equivalent, select BALB/c female mice abdominal cavity and dorsal sc multi-point injection in 6 ~ 8 week age, dosage 0.4mL/ only; 7d, 21d carry out second time, third time immunity respectively, antigen equivalent Fu Shi Freund's incomplete adjuvant emulsification; 35d tail vein injection is without adjuvant antigen 80 μ g, and blood sampling of docking afterwards for three days measures serum ELISA and tires, and chooses the aseptic spleen of getting of higher mouse of tiring and merges.
1.2.2 SP2/0 cell prepares
Merge first 2 weeks recovery SP2/0 myeloma cells, carry out Secondary Culture, after cell growth state is stable, by myeloma cell's enlarged culturing, observation of cell growth conditions, selection is evenly distributed, size is homogeneous, eugonic cell merges.Merge the same day, draw plasma-free DMEM medium with suction pipe and blown down from bottle wall by cell, be collected in centrifuge tube, the centrifugal 5min of 1000r/min, abandons supernatant, myeloma cell is suspended with serum-free DMEM, and trypan blue counting is for subsequent use.
1.2.3 feeder layer cells preparation
After BALB/c mouse after booster immunization being extractd eyeball bloodletting, disconnected cervical vertebra is put to death, and is soaked in 75% alcohol disinfecting 3 ~ 5min.Cut off skin by sterile scissors, expose peritonaeum.Inject 6 ~ 8mL serum-free DMEM nutrient solution with asepsis injector to intraperitoneal, repeatedly rinse 3 ~ 4 times and sucking-off nutrient solution, put into 10mL centrifuge tube, the centrifugal 5 ~ 6min of 1200r/min.Containing 10% top grade foetal calf serum nutrient solution suspendible, packing 96 orifice plate after adjustment cell count 1 × 105/mL, 100 μ L/ holes, the 37 DEG C of constant temperature cell culture incubators put containing 5%CO2 are cultivated.General feeder cell are in fusion preparation the day before yesterday, a mouse can obtain 5 × 106 ~ 8 × 106 peritoneal macrophages, if during with mouse chest cell as feeder cell, cell concn is 5 × 106/mL, mouse boosting cell is 1 × 106/mL, inoblast 1 × 105/mL of mouse, is 100 μ L/ holes.
1.2.4 immune spleen cell
The preparation procedure of splenocyte suspension is as follows: get the BALB/c mouse after booster immunization, as the positive control of antibody test after excision eyeball blood sampling separation of serum.By drawing neck to dislocate lethal mouse, being soaked in 75% alcohol the 3 ~ 5min that sterilizes, with skin on the left of sterile scissors abdominal cut after fixing on autopsy table, cutting off peritonaeum, exposing spleen.Spleen sterile scissors is taken out and is placed in the plate filling the incomplete substratum of 10mL, wipe out surrounding connective tissue, wash gently, put after 200 order copper mesh grind to form cell suspension with syringe nook closing member and count.After general immunity, spleen volume is about 2 times of normal mice spleen volume, and cell count is about 2 × 108.By the centrifugal 5min of splenocyte suspension 1000r/min of results, abandon supernatant, for subsequent use.
1.2.5 cytogamy
Asepticly take immune mouse spleen, splenocyte and SP2/0 cell count respectively, centrifugal after resuspended with incomplete substratum, splenocyte and SP2/0 cell number add in 50mL centrifuge tube with 5:1 ratio, mixing, the centrifugal 5min of room temperature 1000r/min, patting centrifuge tube after discarding incomplete substratum makes cell evenly be attached at the bottom of pipe, slowly drips the PEG3350 solution 1mL that concentration is 50%, rotate centrifuge tube gently in this process under 37 DEG C of water bath condition in 60s; PEG3350 dropwises rear standing centrifuge tube 30s makes cell fully merge, and the incomplete nutrient solution 20mL then dropwise adding 37 DEG C of preheatings from slow to fast in 2min stops merging; Room temperature 800r/min is centrifugal, and 8min abandons supernatant, and appropriate 37 DEG C of preheatings select nutrient solution suspension cell completely containing HAT, joins and cultivates previously prepared containing in 96 porocyte culture plates of feeder cell.Respectively at after fusion the 3rd, 7d HAT Selective agar medium half amount changes liquid, uses HT substratum after 10d instead.Treat that cell colony volume reaches 1/3 culture hole, get supernatant liquor indirect ELISA and carry out antibody test.
1.2.6 indirect ELISA detection method is set up:
(1) bag quilt: after restructuring Bac-VP60a and Bac-VP602 albumen dilutes with pH9.6CBS damping fluid, every hole 200 μ g, coated elisa plate, 4 DEG C are spent the night;
(2) wash: as washings, enzyme plate is washed four times using PBST (the PBS damping fluid of 0.01M, pH7.4 adds the polysorbas20 of 0.05%), every minor tick soaks 3min, and button is dry;
(3) close: the skimming milk PBST solution of 8% (w/v), 250 μ L/ holes, 37 DEG C of closed 2h, the same washing, button are done;
(4) detect: add Hybridoma Cell Culture thing supernatant liquor, 100 μ L/ holes, washing after 37 DEG C of effect 60min, button are done;
(5) add the goat-anti rabbit horseradish peroxidase-labeled IgG of 1:30000 dilution, 100 μ L/ holes, washing after 37 DEG C of effect 1h, button are done;
(6) develop the color: add tmb substrate 100 μ L/ hole, 37 DEG C of lucifuge colour developing 15min;
(7) stop: enzyme plate adds the H2SO4 50 μ L/ hole color development stopping of 2M, microplate reader reads OD490.
The hybridoma cell strain prepared using recombinant protein VP60a as immunogen, for restructuring Bac-VP60a albumen as detectable antigens detected result OD490 value >=2.0 and when negative control hole OD490 value (about 0.2) is equal to the detected result OD490 value using recombinant protein Bac-VP602 as detectable antigens, this hybridoma cell strain, as the sample to be selected identifying separately RHDVa virus, carries out next step subclone and screening.The hybridoma cell strain prepared using recombinant protein VP602 as immunogen, for restructuring Bac-VP602 albumen as detectable antigens detected result OD490 value >=2.0 and when negative control hole OD490 value (about 0.2) is equal to the detected result OD490 value using recombinant protein Bac-VP60a as detectable antigens, this hybridoma cell strain, as the sample to be selected identifying separately RHDV2 C-type virus C, carries out next step subclone and screening.
1.2.7 filtering hybridoma and clone
When cell colony volume grows to 1/3 culture hole (about 10d) time, detect cells and supernatant with indirect ELISA, the hybridoma of screening secretion positive antibody, every hole duplicate detection three times.Gained positive hybridoma cell is carried out three subclones by limiting dilution assay, and limiting dilution assay carries out subclone, and program is as follows:
(1) feeder cell suspension (before merging) is prepared.
(2) positive porocyte counting and adjust cell count at 1 × 103 ~ 5 × 103/mL.
(3) get 130 cells and put into 6.5mL containing feeder cell complete culture solution, i.e. 20/mL, 100 μ L/ holes add A, B, C tri-and arrange as 2, every hole cell.Remaining 2.9mL cell suspension adds the complete culture solution of 2.9mL containing feeder cell, and cell count is 10/mL, and 100 μ L/ holes add D, E, F tri-row, are 1, every hole cell.Remaining 2.2mL cell suspension adds the complete culture solution of 2.2mL containing feeder cell, cell count 5/mL, 100 μ L/ holes, and adding G, H two row, is 0.5, every hole cell.
(4), after cultivating 4 ~ 5d, inverted microscope can see little cell clone, add complete culture solution 100 μ L/ hole.
During (5) 8th ~ 9d, naked eyes visible cell clone (cell colony volume grows to 1/3 culture hole), carries out antibody test in time.The hybridoma of first cloning needs to add HAT in complete culture solution.
(6) clone more than 3 times continuously according to the method described above, until all clone cell holes are all positive.
Can the hybridoma cell strain of stably excreting monoclonal antibody, liquid nitrogen cryopreservation after enlarged culturing.
1.2.8 the Western blot of the antibody molecule of hybridoma secretion identifies
By Bac-VP60a albumen and and Bac-VP602 albumen transferring film after SDS-PAGE electrophoresis, using the culture supernatant of hybridoma cell strain as first antibody, using HRP mark sheep anti-mouse igg as second antibody, DAB substrate develop the color.
1.2.9 the IFA qualification of the antibody molecule of hybridoma secretion
Sf9 cell 72h is infected with rBacmid-VP60a and rBacmid-VP602, abandon cell culture fluid, after paraformaldehyde is fixing, add the culture supernatant of hybridoma cell strain as first antibody, using the sheep anti-mouse igg of FITC mark as second antibody, by the microscopic examination of 488nm wavelength fluorescent.
1.2.10 the hypotype qualification of the antibody molecule of hybridoma secretion
Mouse monoclonal antibody hypotype identification kit, purchased from Yi Qiao Divine Land, Beijing biotech company, is identified the antibody molecule hypotype of hybridoma cell strain secretion to specifications.
Through continuous 3 time clonings, acquisition can secretory antibody molecule identify separately hybridoma cell strain mouse hybridoma cell (Mus musculus Hybridoma) the McAb 1H2 of RHDVa C-type virus C, the antibody molecule of this hybridoma cell strain secretion can identify RHDVa virus particle and restructuring Bac-VP60a albumen, nonrecognition Bac-VP602 albumen, difference extremely remarkable (P<0.01), as shown in Figure 3.Acquisition can secretory antibody molecule identify separately hybridoma cell strain mouse hybridoma cell (Mus musculus Hybridoma) the McAb 1G2 of RHDV2, the antibody molecule of this hybridoma cell strain secretion can not identify RHDVa virus particle and restructuring Bac-VP60a albumen, only identify Bac-VP602 albumen, difference extremely remarkable (P<0.01), as shown in Figure 4.
The antibody molecule that hybridoma cell strain mouse hybridoma cell (Mus musculus Hybridoma) McAb 1H2 secretes can identify restructuring Bac-VP60a albumen, specific band is had at 70kDa place, nonrecognition Bac-VP602 albumen, without band, as shown in Figure 5.The antibody molecule that hybridoma cell strain mouse hybridoma cell (Mus musculus Hybridoma) McAb 1G2 secretes can not identify restructuring Bac-VP60a albumen, only identify Bac-VP602 albumen, specific band is had, as shown in Figure 6 at 70kDa place.
The antibody molecule that hybridoma cell strain mouse hybridoma cell (Mus musculus Hybridoma) McAb 1H2 secretes can identify the Sf9 cell that rBacmid-VP60a infects, the Sf9 cell (as shown in Figure 7) that nonrecognition rBacmid-VP602 infects.The antibody molecule that hybridoma cell strain mouse hybridoma cell (Mus musculus Hybridoma) McAb 1G2 secretes can not identify the Sf9 cell that rBacmid-VP60a infects, and only identifies the Sf9 cell (as shown in Figure 8) that rBacmid-VP602 infects.
The monoclonal antibody of the rabbit haemorrhagic disease RHDV2 C-type virus C that hybridoma cell strain McAb 1G2 of the present invention secretes can identify separately RHDV2 C-type virus C and VP602 albumen; This monoclonal antibody molecule heavy chain is IgG2a hypotype, and light chain is Kappa hypotype (as shown in Figure 2).
And the monoclonal antibody of the rabbit haemorrhagic disease RHDVa C-type virus C that hybridoma cell strain McAb 1H2 secretes can identify separately RHDVa C-type virus C and VP60a albumen; This monoclonal antibody molecule heavy chain is IgG1 hypotype, and light chain is Kappa hypotype (as shown in Figure 1).
Whether what the rabbit that experiment proves to utilize the monoclonal antibody of rabbit haemorrhagic disease RHDV2 C-type virus C of the present invention to identify accurately suffers from rabbit haemorrhagic disease specifically infected is RHDV2 C-type virus C.
Claims (2)
1. hybridoma cell strain McAb 1G2, it is characterized in that it is mouse hybridoma cell (Mus musculusHybridoma) McAb 1G2, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCC No.10316.
2. the monoclonal antibody of rabbit haemorrhagic disease RHDV2, is characterized in that the monoclonal antibody of rabbit haemorrhagic disease RHDV2 C-type virus C, and the hybridoma cell strain McAb 1G2 being CGMCC No.10316 by preserving number produces.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105277696A (en) * | 2015-10-26 | 2016-01-27 | 中国农业科学院上海兽医研究所 | Double-antibody sandwich ELISA (enzyme-linked immuno sorbent assay) detection kit for RHDV (rabbit hemorrhagic disease virus) and application method |
| CN115725511A (en) * | 2022-08-29 | 2023-03-03 | 四川农业大学 | Hybridoma cell strain R2McAb2A1, monoclonal antibody secreted by same and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011135075A1 (en) * | 2010-04-29 | 2011-11-03 | Riemser Arzneimittel Ag | Parapoxvirus expressing the vp60 major capsid protein of the rabbit haemorrhagic disease virus |
| CN102250239A (en) * | 2011-06-13 | 2011-11-23 | 中国农业科学院哈尔滨兽医研究所 | Protein capable of combining with vp60 protein of rabbit hemorrhagic disease virus and use thereof |
| CN102304529A (en) * | 2011-08-31 | 2012-01-04 | 中国农业科学院生物技术研究所 | Preparation method of rabbit hemorrhagic fever virus empty capsid antigen |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011135075A1 (en) * | 2010-04-29 | 2011-11-03 | Riemser Arzneimittel Ag | Parapoxvirus expressing the vp60 major capsid protein of the rabbit haemorrhagic disease virus |
| CN102250239A (en) * | 2011-06-13 | 2011-11-23 | 中国农业科学院哈尔滨兽医研究所 | Protein capable of combining with vp60 protein of rabbit hemorrhagic disease virus and use thereof |
| CN102304529A (en) * | 2011-08-31 | 2012-01-04 | 中国农业科学院生物技术研究所 | Preparation method of rabbit hemorrhagic fever virus empty capsid antigen |
Non-Patent Citations (3)
| Title |
|---|
| GHISLAINE LE GALL-RECULÉ ET AL.: "Emergence of a new lagovirus related to Rabbit Haemorrhagic Disease Virus", 《VETERINARY RESEARCH》 * |
| 刘怀然 等: "兔出血症病毒VP60 基因在昆虫细胞中形成病毒样颗粒及其特性", 《中国比较医学杂志》 * |
| 刘怀然 等: "兔出血症病毒单克隆抗体的制备及鉴定", 《中国兽医科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105277696A (en) * | 2015-10-26 | 2016-01-27 | 中国农业科学院上海兽医研究所 | Double-antibody sandwich ELISA (enzyme-linked immuno sorbent assay) detection kit for RHDV (rabbit hemorrhagic disease virus) and application method |
| CN115725511A (en) * | 2022-08-29 | 2023-03-03 | 四川农业大学 | Hybridoma cell strain R2McAb2A1, monoclonal antibody secreted by same and application |
| CN115725511B (en) * | 2022-08-29 | 2024-02-09 | 四川农业大学 | Hybridoma cell strain R2McAb2A1, monoclonal antibody secreted by same and application thereof |
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