Acetamiprid hapten and preparation method thereof as well as application
A technology of acetamiprid and hapten, which is applied in the field of preparation of acetamiprid hapten and antigen, and achieves high sensitivity, high antibody specificity and rapid detection effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0022] Example 1 Preparation of acetamiprid hapten
[0023] 1. Synthesis of acetamiprid hapten
[0024] Take 0.50 g of N-desmethyl acetamiprid, add 50 mL of methanol to dissolve, add 0.30 g of aminobutyric acid, stir, add 0.37 mL of 37% formaldehyde solution, stir, mix well, react at 80°C for 4 h; stop the reaction, spin evaporate Remove methanol, add 50 mL of water, add 100 mL of ethyl acetate for extraction, extract three times, combine the organic phases, evaporate to dryness, put on a silica gel column, dichloromethane / methanol ( V / V , 10 / 1) was eluted and separated to obtain 0.69 g of the hapten product carboxyacetamiprid, with a yield of 92.81%.
[0025] 2. Identification of acetamiprid hapten
[0026] The above haptens were identified by H NMR spectroscopy, 1 H NMR (CDCl 3 , 300MHZ) δ: 11.0 (1H, -COOH), 8.53 (1H, s, ArH), 7.86 (1H, d, ArH), 7.21 (1H, d, ArH), 4.16 (1H, dd, =CH-) ,3.91 (1H, s, -CH 2 -), 3.21 (2H, d, -CH 2 -), 2.82 (2H, t, -CH 2 -), 2.49 (2H, t...
Embodiment 2
[0028] Example 2 Preparation of Acetamiprid Antigen
[0029] 1. Synthesis of Acetamiprid Immunogen
[0030] The acetamiprid hapten was conjugated with bovine serum albumin (BSA) to obtain the immunogen.
[0031] Take 11 mg of the hapten carboxyacetamiprid, add 1 mL of dimethyl sulfoxide to dissolve, add 0.18 mL of isobutyl chloroformate, and 0.1 mL of triethylamine, and react for 1 h at 0-4°C to obtain Hapten Activation Solution A ; Take 50 mg of BSA, add 3 mL of 0.8% saline to dissolve to obtain liquid B; add liquid A dropwise to liquid B, continue to stir for 5 h, 0.02 mol / L PB dialysis purification for 3 days, change the liquid 3 times a day, The immunogen was obtained, aliquoted, and stored at -20°C.
[0032] 2. Synthesis of acetamiprid coating agent
[0033] The acetamiprid hapten was coupled with ovalbumin (OVA) to obtain the coating source.
[0034] Take 5 mg of the hapten carboxyacetamiprid, add 1 mL of dimethyl sulfoxide to dissolve, add 9 mg of dicyclohexylcarbod...
Embodiment 3
[0037] Example 3 Preparation of Acetamiprid Monoclonal Antibody
[0038] 1. Obtaining hybridoma cells
[0039] 1) First immunization: Fully emulsify the acetamiprid hapten-BSA conjugate (immunogen) with an equal amount of Freund's complete adjuvant, and inject 6-week-old Balb / c mice subcutaneously, 0.2 mL each;
[0040] 2) Booster immunization twice: from the first immunization, booster immunization once every two weeks, with Freund's incomplete adjuvant instead of Freund's complete adjuvant, the method and dosage are the same as the first immunization;
[0041] 3) One week after the last booster immunization, the fundus vein blood was collected to measure the titer and inhibition. When there was inhibition and the titer reached more than 1:10000, the following last immunization was carried out: intraperitoneal injection of 0.1 mL of the immunogen solution without any adjuvant, and then executed three days later Mice, whose spleen was fused with myeloma cells;
[0042] 4) Us...
PUM
Login to view more 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