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Preparation method and application of forchlorfenuron artificial antigen and polyclonal antibody

A polyclonal antibody and artificial antigen technology, applied in chemical instruments and methods, animal/human protein, serum albumin, etc., can solve the problems of unsuitable for large-scale on-site screening, cumbersome and complicated operation, high detection cost, etc. Effects of radioactive isotope contamination, simple operation, and high sensitivity

Inactive Publication Date: 2018-01-23
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Forchlorfenuron residue detection currently mainly adopts liquid chromatography, gas chromatography or mass spectrometry, which has problems such as high detection cost, long cycle and cumbersome and complicated operation, and is not suitable for large-scale on-site screening. Therefore, it is urgent to develop simple , sensitive, specific, and low-cost rapid detection technology

Method used

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  • Preparation method and application of forchlorfenuron artificial antigen and polyclonal antibody
  • Preparation method and application of forchlorfenuron artificial antigen and polyclonal antibody
  • Preparation method and application of forchlorfenuron artificial antigen and polyclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0036] Embodiment: a kind of preparation method of forchlorfenuron artificial antigen and polyclonal antibody, concrete steps are as follows:

[0037] (1) Synthesis of forchlorfenuron hapten

[0038] Using anhydrous dimethylformamide (DMF) as solvent, forchlorfenuron (2.48 g), succinic anhydride (1 g), and anhydrous aluminum trichloride (6.68 g) were carried out at a molar ratio of 1:1:5. Feeding. Add DMF (2 mL) dropwise into the flask containing anhydrous aluminum trichloride, stir continuously to dissolve it fully, and put it into an oil bath (200°C) when there is no white mist coming out of the bottle mouth. Add the mixture of forchlorfenuron and succinic anhydride into the flask, and after reacting for 2 h, slowly pour the reaction liquid into 90 mL of ice water, add 6 mL of concentrated hydrochloric acid with a concentration of 12 mol / L, let it cool and stand, and then filter Wash with water to obtain the crude product, and then recrystallize and purify the crude produc...

experiment example 3

[0071] Experimental Example 3: Antibody Specificity Experiment

[0072] Select diuron, rituron, chlorotoluron, picloram, and clopyralid, which are similar in structure to forchlorfenuron, to prepare a series of standard solutions with different concentrations to replace the forchlorfenuron standard solution, determine its standard curve, and Calculation IC 50 The inhibitory concentration was used to obtain the cross-reactivity rate (CR (%)). The results are shown in Table 4.

[0073] CR (%) = (IC 50 Concentration of forchlorfenuron / IC 50 Concentration of analogue at time) × 100%

[0074] Table 4 Cross-reactivity of forchlorfenuron and its analogues

[0075]

[0076] The results showed that the polyclonal antibody against forchlorfenuron hardly combined with other analogues, and the cross-reactivity rate was very small. It shows that the antibody has high specificity to forchlorfenuron.

[0077] Utilizing the indirect competitive ELISA method of the present invention t...

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Abstract

The invention discloses a preparation method and application of a forchlorfenuron artificial antigen and polyclonal antibody. The preparation method mainly includes the steps of firstly, using Friedel-Crafts reaction to synthesize artificial forchlorfenuron hapten; secondly, using the artificial forchlorfenuron hapten as the raw material, and using a carbodiimide method to couple the artificial forchlorfenuron hapten with bovine serum albumin to obtain the forchlorfenuron artificial antigen; thirdly using the synthesized artificial antigen to immunize New Zealand white rabbits, taking blood, centrifuging to obtain antiserum, namely the anti-forchlorfenuron polyclonal antibody. The preparation method and the application have the advantages that the problems that a traditional physicochemical analysis method is complex, high in cost, slow in analysis and the like are solved, and the immunoassay technology which is simple, fast and sensitive is provided; the prepared antibody is good in specificity and sensitivity, simple in synthesizing method, applicable to the fast immune detection of forchlorfenuron and good in application value.

Description

technical field [0001] The invention relates to a preparation method and application of forchlorfenuron artificial antigen and polyclonal antibody, and belongs to the fields of plant growth regulator small molecular compound immunochemistry and food safety analysis. Background technique [0002] Forchlorfenuron is a highly active plant growth regulator, which can accelerate cell mitosis, promote cell growth and differentiation, prevent flower drop and fruit drop, and promote fruit expansion. It is often used in navel oranges, citrus, kiwi, grapes, watermelons, and cucumber In the agricultural production of fruits and vegetables. Forchlorfenuron residues in fruits and vegetables may pose a potential threat to humans and other organisms. The US Environmental Protection Agency pointed out in "Pesticide Toxicity Confirmation and Management" that long-term exposure to forchlorfenuron can cause protein metabolism in the body. Swelling and emaciation. Different countries set diff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C07K16/44G01N33/535G01N33/543
Inventor 郭岚万益群肖石妹鄢爱平柳英霞
Owner NANCHANG UNIV
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