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Trinexapac-ethyl artificial antigens, a preparing method of a specific antibody and uses of the antibody

A technology for trinexapac-ethyl and hapten, which is applied in the field of immunochemical analysis to achieve the effects of low cost, convenient on-site monitoring and strong antibody specificity

Inactive Publication Date: 2017-12-15
NINGBO ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on the artificial haptens, artificial antigens and antibodies of trinexapac-ethyl at home and abroad

Method used

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  • Trinexapac-ethyl artificial antigens, a preparing method of a specific antibody and uses of the antibody
  • Trinexapac-ethyl artificial antigens, a preparing method of a specific antibody and uses of the antibody
  • Trinexapac-ethyl artificial antigens, a preparing method of a specific antibody and uses of the antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Synthesis of artificial haptens

[0035] Carboxylate on the six-membered ring of trinexapac-ethyl is subjected to alkaline hydrolysis to synthesize a hapten, which not only retains the characteristic group of trinexapac-ethyl to the greatest extent, but also forms a carboxyl terminal group, which can be better coupled with the protein. After purification, the product was identified by mass spectrometry (ESI) and hydrogen nuclear magnetic resonance spectrum (1H-NMR). The molecular structural formula of trinexapac-ethyl artificial hapten is:

[0036]

[0037] The specific preparation method is as follows: Weigh 5.04g (about 0.02mol) of trinexapac-ethyl and 30mL of tetrahydrofuran into a 100mL conical flask, then add 24mL of 5% sodium hydroxide methanol solution (trinexapac-ethyl: sodium hydroxide=1: 1.5 mol ratio), stirred at room temperature for 4 h and added 20 mL of water; the mixed solution was extracted with ethyl acetate, the organic phase was discarded, the ...

Embodiment 2

[0039] Identical to the preparation method of Example 1, the difference is that the proportioning of each raw material is different:

[0040] The specific preparation method is as follows: Weigh 7.56g (about 0.03mol) of trinexapac-ethyl and 40mL of tetrahydrofuran into a 100mL conical flask, then add 36mL of 5% sodium hydroxide methanol solution (trinexapac-ethyl: sodium hydroxide=1: 1.5 mol ratio), stirred and reacted at room temperature for 4 h and added 30 mL of water; the mixed solution was extracted with ethyl acetate, the organic phase was discarded, the aqueous phase was adjusted to pH 2.0 with concentrated hydrochloric acid, extracted with ethyl acetate, and the organic phases were combined, without After drying over sodium sulfate, it was concentrated under reduced pressure to obtain a yellow oil; it was purified with a silica gel column, and the eluent was a mixture of 150 mL of methanol:dichloromethane=1:40 (volume ratio), and the fractions were collected and concent...

Embodiment 3

[0042] Identical to the preparation method of Example 1, the difference is that the proportioning of each raw material is different:

[0043]The specific preparation method is as follows: Weigh 10.08g (about 0.04mol) of trinexapac-ethyl and 60mL of tetrahydrofuran into a 100mL conical flask, then add 48mL of 5% sodium hydroxide methanol solution (trinexapac-ethyl: sodium hydroxide=1: 1.5 molar ratio), stirred at room temperature for 5 h and added 40 mL of water; the mixed solution was extracted with ethyl acetate, the organic phase was discarded, the aqueous phase was adjusted to pH 2.0 with concentrated hydrochloric acid, extracted with ethyl acetate, and the organic phases were combined, without After drying with sodium sulfate, it was concentrated under reduced pressure to obtain a yellow oil; it was purified with a silica gel column, and the eluent was a mixture of 110 mL of methanol:dichloromethane=1:50 (volume ratio), and the fractions were collected and concentrated to n...

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Abstract

The invention discloses trinexapac-ethyl antigens, a specific antibody and uses of the antibody, and belongs to the technical field of immunochemical analysis. The antigens are specially used for preparation of the trinexapac-ethyl specific antibody and an enzyme-linked immunological detection method. The artificial antigens which are TNc-BSA and TNc-OVA are synthesized, a rabbit is immunized to obtain the trinexapac-ethyl specific polyclonal antibody, and finally an enzyme-linked immunosorbent assay method is established by utilizing the synthesized coated antigens and the specific antibody. The lowest trinexapac-ethyl detection limit of the method is 0.005 mg / L, the recovery rate is 85.2-103.1%, and the average coefficient of variability is 2.1-7.9% antibody, thus meeting residue analysis standards.

Description

technical field [0001] The invention belongs to the technical field of immunochemical analysis, and in particular relates to a preparation method and application of an antigen and a specific antibody of a cyclohexane carboxylic acid plant growth regulator. Background technique [0002] Trinexapac-ethyl, the chemical name is 4-cyclopropyl (hydroxyl) methylene-3,5-dioxocyclohexane ethyl carboxylate, and its structure is as follows: [0003] [0004] It belongs to cyclohexane carboxylic acid plant growth regulator, and is a gibberellin biosynthesis inhibitor, which controls plant growth by reducing the content of gibberellin. Trinexapac-ethyl can be rapidly absorbed by plant stems and leaves, but little absorbed by roots. Applied on the leaves, it can be transferred to the growing branches to reduce the elongation of the internodes, and can be used on various crops such as cereal crops, wheat, and sugarcane. It overcomes the shortcomings of many other plant growth regulato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C62/38C07C51/47C07C51/48C07C27/02C07K14/765C07K14/77C07K1/107C07K16/44G01N33/531
CPCC07C51/09C07C51/47C07C51/48C07C62/38C07K14/765C07K14/77C07K16/44C07K19/00G01N33/531
Inventor 付岩张亮张艳陈国吴银良
Owner NINGBO ACAD OF AGRI SCI
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