Preparing method for paclobutrazol monoclonal antibodies

A technology for cloning antibodies and paclobutrazol monoclonal antibodies is applied in the field of preparation of paclobutrazol monoclonal antibodies, and can solve the problems of complicated and complicated operation process, high cost, expensive detection instruments and the like.

Inactive Publication Date: 2015-02-18
NANCHANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Traditional paclobutrazol residue analysis methods mainly rely on instrumental analysis methods such as liquid chromatography, gas chromatography or mass spectrometry, and the detection instruments are expensive. Although these detection

Method used

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  • Preparing method for paclobutrazol monoclonal antibodies
  • Preparing method for paclobutrazol monoclonal antibodies

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Embodiment 1

[0105] A preparation method of paclobutrazol monoclonal antibody, the specific operation method is as follows:

[0106] (1) Synthesis of paclobutrazol hapten

[0107] Using anhydrous pyridine as a solvent, mix 1.1 mg of paclobutrazol, 5-10 mg of succinic anhydride and 0.46 mg of 4-dimethylaminopyridine in a glass centrifuge tube. After mixing evenly, blow dry the solvent with nitrogen, and place the glass tube in a microwave oven. The reaction was carried out intermittently at 406 W for 185 min under medium heat. After the reaction was completed, 1.5 mL of chloroform was added to dissolve the mixture. The mixture was taken out and centrifuged to remove the precipitation. The supernatant was added with 1.5 mL of secondary water, and mixed on a mini shaker for 1-2 min. Chloroform phase, after drying with nitrogen, a pale yellow powder is obtained, which is the hapten paclobutrazol hemisuccinate;

[0108] (2) Synthesis of paclobutrazol immune antigen

[0109] 2 mg and 5 μmoL of...

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Abstract

The invention discloses a preparing method for paclobutrazol monoclonal antibodies. The preparing method comprises the main steps as follows: (1) synthesizing artificial hapten paclobutrazol hemisuccinates by a microwave solvent free method; (2) performing coupling to obtain artificial antigens of the paclobutrazol by using the paclobutrazol hemisuccinates as raw materials; (3) performing the immune treatment on Balb/c mice by using the synthesized artificial antigens; (4) selecting the mice with optimal serum valence and optimal specificity, and mixing spleen cells and myeloma cells of the mice together in an external manner; (5) culturing and screening the fused cells by using the selective culture medium, and further cloning, propagating and storing by freezing; (7) injecting the expanded cell strains into the abdominal cavities of the mice to generate a lot of abdominal dropsy; (8) purifying the paclobutrazol-resistant monoclonal antibodies in the abdominal dropsy by a caprylic acid-saturated ammonium sulfate method. The prepared paclobutrazol-resistant monoclonal antibodies have high sensitivity and strong specificity, and can be further applied to the construction of technologies of immune sensors, colloidal gold immune chromatographic methods and the like.

Description

[0001] technical field [0002] The invention relates to a preparation method of paclobutrazol monoclonal antibody, belonging to the technical field of monoclonal antibody preparation. Background technique [0003] Paclobutrazol is a growth-inhibiting plant growth regulator. It has the effects of delaying plant growth, inhibiting stem elongation, shortening internodes, promoting flower bud differentiation, increasing plant stress resistance, and increasing yield. Therefore, it is widely used in rice, Wheat, peanuts, fruit trees, tobacco, rapeseed, soybeans, flowers, lawns, etc. as (plants). With the continuous expansion of the use of plant growth regulators, the chronic and long-term effects of their residues on the environment and human health have increasingly aroused people's attention and concern. For this reason, the restrictions on the residues of plant growth regulators are becoming more and more strict. my country has stricter limit standards for the residues of pac...

Claims

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Application Information

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IPC IPC(8): C07K16/44
Inventor 万益群郭岚刘香香柳英霞鄢爱平谭婷
Owner NANCHANG UNIV
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