Monoclonal antibody of anti lymphocyst virus of and preparation method
A technology of lymphocyst virus and monoclonal antibody, which is applied in the cross field of molecular immunology and virology, to achieve the effect of good adhesion, uniform cell shape and strong refraction
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0037] Development of monoclonal antibodies against flounder lymphocyst virus:
[0038] 1. Purification of flounder lymphocyst virus:
[0039] (1) Take the diseased flounder, first use 70% alcohol cotton ball to disinfect the affected part, then cut off the cyst with a sterilized scalpel, add an appropriate amount of quartz sand and 10 times the volume of TNE buffer (100mM NaCl, 1mM EDTA, 50mM Tris , pH 7.4), homogenate;
[0040] (2) The homogenate was centrifuged at 4°C, 500g, for 30min, and the supernatant was taken;
[0041] (3) The supernatant was centrifuged at 4°C, 1800g, for 30min, and the supernatant was taken again;
[0042] (4) Prepare 30% (W / V) sucrose solution with sucrose and supernatant, centrifuge at 78500g for 120min at 4°C, discard supernatant;
[0043] (5) Add TNE to the precipitate to 1ml, place it lightly above the sucrose gradient (37%, 40%, 47%, 52%, 57%, 62%) (W / V), centrifuge at 78500g for 120min at 4°C ;
[0044] (6) Carefully suck out the virus b...
Embodiment 2
[0081] Identification of flounder lymphocyst virus monoclonal antibody of the present invention:
[0082] 1. Identification by indirect immunofluorescence antibody method:
[0083] First, the production of frozen sections:
[0084] Fresh cysts were taken, cut into small pieces about 3mm square, washed with normal saline, blotted dry, embedded with frozen embedding agent, placed at -20°C for 30min, and sliced at -20°C with a cryostat, with a thickness of 5 μm. The slices were fixed with acetone for 20 min, dried at room temperature, and frozen at -20°C for later use.
[0085] Second, fluorescent antibody staining:
[0086] (1) The culture supernatant of the hybridoma cells screened and cloned above was used as the primary antibody, and added to the slice.
[0087] (2) Incubate in a humid box at 37°C for 45 minutes.
[0088] (3) Take out the glass slide and wash it 3 times with 0.01M phosphate buffered saline, 5 min each time.
[0089] (4) Goat anti-mouse IgG antibody lab...
Embodiment 3
[0110] Using the developed monoclonal antibody against flounder lymphocyst virus to identify Sebastesschlegeli lymphocyst virus;
[0111] First, the production of frozen sections:
[0112] Take the diseased flat scorpionfish cyst tissue, cut into small pieces about 3 mm square, wash with normal saline, blot dry, embed with frozen embedding agent, place at -20°C for 30min, and slice at -20°C with a frozen microtome. The slice thickness is 5 μm. The slices were fixed with acetone for 20 min, dried at room temperature, and frozen at -20°C for later use;
[0113] Second, fluorescent antibody staining:
[0114] (1) Aspirate 15 μl of monoclonal antibody against flounder lymphocyst virus and add it to the slice.
[0115] (2) Incubate in a humid box at 37°C for 45 minutes;
[0116] (3) Take out the glass slide and wash it 3 times with 0.01M phosphate buffered saline, 5 min each time.
[0117] (4) Goat anti-mouse IgG antibody labeled with fluorescein isothiocyanate, added to the s...
PUM
| Property | Measurement | Unit |
|---|---|---|
| thickness | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to view more - R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap