Immunomagnetic beads enriched with t-2 toxin and its preparation method and application
An immunomagnetic bead and toxin technology, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of low separation efficiency, complicated purification and separation of T-2 toxin samples, etc., to improve the detection limit, improve detection accuracy and Reliability, the effect of eliminating the interference of impurities
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0029] Example 1 Preparation of immunomagnetic beads enriched for T-2 toxin
[0030] This example provides a method for preparing the conjugate obtained by coupling the T-2 toxin monoclonal antibody with carboxyl-containing immunomagnetic beads as the immunomagnetic beads enriched in T-2 toxin. The method includes:
[0031] 1. Preparation of T-2 toxin monoclonal antibody
[0032] 1. Synthesis of T-2 toxin hapten (synthetic route see appendix figure 1 ) and identification
[0033] Dissolve 100mg of T-2 toxin in acetonitrile, then add 200μL of acetic acid and 200mg of pyridinium dichromate, react at 70°C for 4 hours, stop the reaction, rotary evaporate, remove acetonitrile, add water-ethyl acetate extraction, dry, evaporate to dryness, and apply silica gel The column was separated by elution with chloroform / methanol (10:1) to obtain an intermediate product.
[0034] Add 3 mL of ethanol to dissolve the above intermediate product, then add 50 mg of potassium carbonate and 5 mL...
Embodiment 2
[0065] Example 2 Characteristic Detection of Immunomagnetic Beads
[0066] Take 0.1 mL of T-2 toxin-enriched immunomagnetic beads prepared according to Example 1 (concentration: 10 mg / mL) in a 10 mL centrifuge tube, wash the magnetic beads twice with 5 mL of deionized water, and remove the upper beads after magnetic separation. clear; then add 1mL of the sample to be tested (the T-2 toxin standard was prepared with PBS buffer solution to T -2 toxin solution as the sample to be tested, and PBS buffer as the blank sample to be tested), mix well, capture at 25°C for 20 minutes, and mix the magnetic beads for 5 minutes during the period; remove the supernatant after magnetic separation, and wash with 5mL deionized water Rinse the magnetic beads twice to remove interfering impurities. Finally, 1 mL of methanol was added for elution, the eluate was collected, and the content of T-2 toxin in the sample to be tested was detected by HPLC according to GB / T28718-2012. The results are s...
Embodiment 3
[0070] The usage method of embodiment 3 immunomagnetic beads
[0071] 1. Sample pretreatment
[0072] Grain and feed samples: Homogenize the sample with a homogenizer; weigh 5g (accurate to 0.01g) of the sample into a sample bottle, add 1.5g of sodium chloride and 30mL of 60% methanol solution, and vortex with a vortex for 5min, or Shake for 20 minutes, over 3000g, and centrifuge at room temperature (20-25°C / 68-77°F) for 5 minutes; absorb 5mL of centrifuged supernatant, add 5mL of deionized water, mix well, and set aside.
[0073] 2. Immunomagnetic bead capture
[0074]Take 0.2 mL of T-2 toxin immunomagnetic beads in a 10 mL centrifuge tube, wash them twice with 5 mL of deionized water, and separate the washing solution with a magnetic separation rack each time (stand still on the magnetic separation rack for 3 minutes each time to ensure that the magnetic beads All adsorption); Add 5mL of the processed sample to the rinsed T-2 toxin immunomagnetic beads, mix well, capture a...
PUM
Property | Measurement | Unit |
---|---|---|
particle diameter | aaaaa | aaaaa |
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