Paraquat hapten, paraquat complete antigen and preparation method of paraquat hapten and paraquat complete antigen

A technology of complete antigen and paraquat, which is applied to the preparation method of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problems of complex processing, cumbersome operation, and poor measurement accuracy, and achieve simple and effective synthesis steps. Good specificity and low cost effect

Inactive Publication Date: 2013-06-05
INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the accurate determination methods of paraquat include gas chromatography (GC), high performance liquid chromatography (HPLC), mass spectrometry, high performance capillary electrophoresis (HPCE), thin layer chromatography (TLC), etc., although TLC is easy to operate , but the operator must directly contact the standard during the detection process, the measurement accuracy is poor, and the detection sensitivity is not high, so it can only be used for semi-quantitative analysis
GC, HPLC and mass spectrometry can be used for accurate quantitative analysis of paraquat in samples, but the sample pretreatment process is complicated, the operation is cumbersome, and all require expensive equipment and professional operators, which is not suitable for rapid on-site detection

Method used

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  • Paraquat hapten, paraquat complete antigen and preparation method of paraquat hapten and paraquat complete antigen
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  • Paraquat hapten, paraquat complete antigen and preparation method of paraquat hapten and paraquat complete antigen

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Embodiment 1

[0032] Example 1, Preparation and identification of paraquat hapten

[0033] 1. Preparation of Paraquat Hapten

[0034] The synthesis route of paraquat hapten is as follows:

[0035]

[0036] Specific synthesis process:

[0037] (1) Synthesis of N-methyl-dipyridine iodide A 250ml three-necked round-bottomed flask was fixed on an iron stand, protected from light and passed through nitrogen. 808 mg of 4,4'-bipyridine was dissolved in 8 ml of acetone, stirred and cooled to 4°C, and 400 μl of methyl iodide was added dropwise to the above solution. Keep stirring at 4°C. The end point of the reaction was controlled by TLC (developing solvent: methanol: glacial acetic acid = 5:2). The reaction reached equilibrium in about 6 hours. Filter, wash with acetone, and drain to obtain yellow needle-like crystals. The crystals were recrystallized from absolute ethanol. Vacuum dry.

[0038] (2) Synthesis of N-methyl-N'-pentanoic acid ethyl ester group-dipyridine iodide bromide 0.85m...

Embodiment 2

[0042] Example 2, Preparation and Identification of Paraquat Complete Antigen

[0043] 1. Preparation of paraquat complete antigen

[0044] The synthetic route of paraquat complete antigen is as follows:

[0045]

[0046]Specific synthesis process: Dissolve 9 mg of paraquat hapten in 500 ul of dimethyl sulfoxide (DMSO), add 15 ul of tri-n-butylamine, stir at room temperature for 5 min, then add 8 ul of isobutyl chloroformate, and stir at room temperature for 30 min. 15 mg of BSA was dissolved in 2 ml of borate buffer solution pH 8.7. Slowly drop the paraquat hapten solution into the BSA solution. Stir overnight at 4°C, dialyze with PBS for three days, and change the dialysate every four hours. Aliquot into EP tubes and store at -20°C.

[0047] 2. Identification of paraquat complete antigen

[0048] According to the increase of the molecular weight of the complete antigen after coupling compared to the molecular weight of the carrier protein, the coupling result was ide...

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Abstract

The invention discloses paraquat hapten, paraquat complete antigen and a preparation method of the paraquat hapten and the paraquat complete antigen. Midbody 4,4-bipyridine of paraquat is utilized to combined analogue N-methyl-N'-valeric acid group-bipyridine-bibromide, carrier protein bovine serum albumin (BSA) is coupled through a mixed anhydride method, and paraquat artificial complete antigen can be successfully combined after ultraviolet spectrum identification and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) identification. The preparation method is simple and effective and low in cost. The combined paraquat artificial complete antigen can be applied to preparing peculiarly identifying efficient antibody of chemical pesticide paraquat and developing immune quick test kits remained by quick detection of paraquat to provide convenient methods for detection research of the materials.

Description

technical field [0001] The invention relates to a paraquat hapten, a paraquat complete antigen, a preparation method of a paraquat hapten and a preparation method of a paraquat complete antigen, belonging to the technical field of biochemical industry, and is used for the preparation and development of an efficient antibody for specific recognition of the chemical pesticide paraquat An immune rapid test kit that can quickly detect paraquat residues. Background technique [0002] Paraquat (Paraquat, PQ), also known as Gramoxone and Paraquat, has a chemical active ingredient of 1,1,-dimethyl-4,4,-bipyridine cationic salt, because of its broad-spectrum, quick-acting, safe, etc. It is widely used in garden weeding and non-cultivated land chemical weeding, and can kill most grasses and broad-leaved weeds. Since the British ICI Crop Protection Company discovered paraquat in 1955, its commercial production scale has expanded year by year. It has been popularized and applied in more ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D213/22C07D213/127C07K14/765C07K14/77C07K14/795C07K1/113
Inventor 宁保安高志贤刘建青彭媛柳明白家磊孙思明
Owner INST OF HYGIENE & ENVIRONMENTAL MEDICINE PLA ACAD OF MILITARY MEDICAL
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