Monoclonal antibody, enzyme-linked immunosorbent assay method and kit for detecting furaltadone residue marker AMOZ
An enzyme-linked immunosorbent reagent, furaltadone technology, applied in the field of detection analysis and immunology, can solve the problems of low efficiency and inability to be widely used, and achieve the effect of high detection efficiency, good accuracy, and simple synthesis process
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0030] Example 1: Synthesis of Antigens
[0031] 1.1 Synthesis of the hapten 3-p-formylbenzoyl-5morpholinomethyl-2-oxazolidinone (CPAMOZ)
[0032] Add 37.5 mg of p-aldehyde benzoic acid into a brown vial, slowly add methanol until completely dissolved, and stir magnetically. Add 50 mg of furaltadone residual marker AMOZ, and react with magnetic stirring at room temperature for 48 hours in the dark. Centrifuge at 8000 rpm for 10 minutes to remove the supernatant, wash with methanol three times, remove the supernatant, and dry the precipitate to obtain the white product CPAMOZ.
[0033]
[0034] 1.2 Synthesis of immunogen (CPAMOZ-BSA conjugate)
[0035] Take CPAMOZ 0.0167g and dissolve in 1ml N,N-dimethylformamide (DMF), add N,N'dicyclohexylcarbodiimide (DCC) 0.0137g and N-hydroxysuccinimide (NHS) 0.0072 g, stirred overnight at 4°C with magnetic force. Centrifuge at 5000rpm for 10min at 4°C and take the supernatant as solution A for later use. Weigh 0.070g of BSA and dis...
Embodiment 2
[0041] Embodiment 2: Preparation of monoclonal antibody
[0042] Preparation of hybridoma cell lines: refer to the method in Xue Qingshan's "Principles and Techniques of In Vitro Culture" Science Press, 2001 edition: immunize Balb / C mice with the CPAMOZ-BSA conjugate prepared in Example 1, and the immunization procedure is: basic Immunization After emulsifying the immunogen with an equal volume of complete Freund's adjuvant, inject it subcutaneously at multiple points on the back of the mouse, and then boost the immunization once every 2 weeks, replace it with incomplete adjuvant emulsification, and finally inject intraperitoneally three days before the fusion , Enhanced immunization, double the amount of antigen, no adjuvant. At the time of fusion, a Balb / C mouse that received the final booster immunization was taken, sacrificed by bloodletting from the orbit (serum was collected, it was positive serum), and sterilized by immersing in 75% alcohol by volume for 5 minutes.
[...
Embodiment 3
[0048] Embodiment 3: the establishment of ELISA detection method
[0049] 3.1 Optimization of ELISA conditions
[0050] Accurately pipette 100 μL of the diluted CPAMOZ-OVA coating original working solution into each well, pack one line for each concentration, and store in the refrigerator at 4°C overnight. Take out the ELISA plate, add 250 μL of washing solution (PBST) to each well, vibrate on the shaker for 1 min, shake off the washing solution, and pat dry on absorbent paper (or clean towel). Repeat the wash 2 times. Accurately pipette 250 μL of blocking solution (1% OVA) into each well, and incubate in a 37°C incubator for 2 hours. Shake off blocking solution and pat dry on absorbent paper. Accurately pipette 250 μL of washing solution into each well, vibrate on a shaker for 1 min, shake off the washing solution, and pat dry on absorbent paper. Repeat the wash 2 times.
[0051] Take the coated ELISA plate, add 50 μL of 20 mM PBS buffer solution with pH 7.4 to each well...
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