Monoclonal antibody hybridoma cell line for paraquat and application of monoclonal antibody hybridoma cell line
A technology of monoclonal antibody and paraquat, which is applied in the application, fusion of cells, and cells modified by introducing foreign genetic material, etc. It can solve the problems of complex pretreatment, inapplicability of rapid detection of a large number of samples, time-consuming, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] Example 1 Preparation of Hybridoma Cell Line LH1
[0033] 1. Derivation of hapten: 4,4'-bipyridine and methyl iodide are the starting materials to synthesize N-methyl-N'-valeryl-dipyridine dibromide as the hapten of paraquat. It is Para-COOH.
[0034] 2. Preparation of complete antigen Para-COOH-KLH: Weigh 2.7mg Para-COOH, 2.7N-hydroxysuccinimide (NHS), and dissolve in 300μL anhydrous N,N-dimethylformamide (DMF) Stir the reaction at room temperature for 10min; then weigh 4.8mg N,N'-dicyclohexylcarbodiimide (DCC), dissolve it with 100μL of anhydrous DMF, add it to the Para-COOH solution, stir at room temperature for 6-8h ( Called A liquid). Take 6.8mg KLH, add 1mL 0.01M phosphate buffer solution (PBS, pH=7.4) to dilute (called B solution), then slowly add A solution to B solution drop by drop, react overnight at room temperature; then use 0.01M PBS solution Dialysis, remove the unreacted small molecule hapten, obtain the complete antigen Para-COOH-KLH, and identify it by u...
Embodiment 2
[0043] (1) Coating: Dilute the coated original Para-COOH-OVA with 0.05M pH9.6 carbonate buffer from 1μg / mL at a 3-fold ratio, 100μL / well, and react at 37°C for 2h.
[0044] (2) Washing: Pour out the solution in the plate and wash it with washing liquid 3 times, 3 minutes each time.
[0045] (3) Blocking: After patted dry, add 200μL / well blocking solution and react at 37°C for 2h. Dry it for later use after washing.
[0046] (4) Adding samples: Dilute the antiserum from 1:1000 in multiples and add it to the coated wells of each dilution, 100μL / well, react at 37°C for 30min; after thorough washing, add 1:3000 diluted HRP -Goat anti-mouse IgG, 100μL / well, react at 37°C for 30min.
[0047] (5) Color development: Take out the ELISA plate and wash it thoroughly, add 100μL of TMB color development solution to each well, and react for 15 minutes at 37°C in the dark.
[0048] (6) Termination and determination: Add 50μL of termination solution to each well to terminate the reaction, and then me...
Embodiment 3
[0050] Take water resources as an example to test the residual amount of paraquat
[0051] (1) Sample pretreatment: Obtain negative samples (named sample 1 and sample 2) from Jiangsu Import and Export Inspection and Quarantine Bureau, add Para and centrifuge, and take 50μL of supernatant (or make a certain dilution) for testing.
[0052] (2) Sample detection: According to the steps of Example 2, the label pull and the actual detection are carried out at the same time. The results are shown in Table 1. The recovery rate is 83.4%-110.9%, the matrix interference is small, and the result is ideal.
[0053] Table 1. ic-ELISA test spiked water samples (n=6)
[0054]
PUM
Abstract
Description
Claims
Application Information
- 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, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com